LOCUS Exported 9565 bp ds-DNA circular SYN 30-AUG-2016
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS Untitled 2
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 9565)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Tuesday, August 30, 2016 from SnapGene Viewer 3.1.4
http://www.miaolingbio.com
FEATURES Location/Qualifiers
source 1..9565
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind complement(10..26)
/note="M13 fwd"
/note="common sequencing primer, one of multiple similar
variants"
promoter 500..604
/gene="bla"
/note="AmpR promoter"
CDS 605..1465
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/note="AmpR"
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
rep_origin 1636..2224
/direction=RIGHT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
protein_bind 2512..2533
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 2548..2578
/note="lac promoter"
/note="promoter for the E. coli lac operon"
protein_bind 2586..2602
/bound_moiety="lac repressor encoded by lacI"
/note="lac operator"
/note="The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-beta-D-thiogalactopyranoside (IPTG)."
primer_bind 2610..2626
/note="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
rep_origin 2664..2799
/note="SV40 ori"
/note="SV40 origin of replication"
polyA_signal 2818..2939
/note="SV40 poly(A) signal"
/note="SV40 polyadenylation signal"
LTR 3011..3244
/note="3' LTR (Delta-U3)"
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
misc_feature 3318..3906
/note="WPRE"
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
CDS 3424..3435
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/note="Factor Xa site"
/translation="IEGR"
CDS complement(3913..5343)
/codon_start=1
/product="tandem dimeric (pseudo-monomeric) derivative of
DsRed (Shaner et al., 2004)"
/note="tdTomato"
/note="mammalian codon-optimized"
/translation="MVSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTA
KLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
LVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH
HLFLGHGTGSTGSGSSGTASSEDNNMAVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPY
EGTQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVM
NFEDGGLVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGV
LKGEIHQALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYE
RSEGRHHLFLYGMDELYK"
CDS complement(5353..5406)
/codon_start=1
/product="2A peptide from Thosea asigna virus capsid
protein"
/note="T2A"
/note="Eukaryotic ribosomes fail to insert a peptide bond
between the Gly and Pro residues, yielding separate
polypeptides."
/translation="EGRGSLLTCGDVEENPGP"
CDS complement(5419..7068)
/codon_start=1
/product="firefly luciferase"
/note="luciferase"
/note="synthetic luc2 version of the luciferase gene"
/translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
LTR 7108..7376
/note="5' LTR (truncated)"
/note="truncated 5' long terminal repeat (LTR) from human
T-cell leukemia virus (HTLV) type 1"
promoter complement(7389..7600)
/note="EF-1-alpha core promoter"
/note="core promoter for human elongation factor
EF-1-alpha"
misc_feature 7645..7762
/note="cPPT/CTS"
/note="central polypurine tract and central termination
sequence of HIV-1"
misc_feature 8257..8490
/note="RRE"
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
misc_feature 8983..9108
/note="HIV-1 Psi"
/note="packaging signal of human immunodeficiency virus
type 1"
LTR 9152..9332
/note="5' LTR (truncated)"
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
ORIGIN
1 cagcttggca ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca
61 acttaatcgc cttgcagcac atcccccttt cgccagctgg cgtaatagcg aagaggcccg
121 caccgatcgc ccttcccaac agttgcgcag cctgaatggc gaatggcgcc tgatgcggta
181 ttttctcctt acgcatctgt gcggtatttc acaccgcata tggtgcactc tcagtacaat
241 ctgctctgat gccgcatagt taagccagcc ccgacacccg ccaacacccg ctgacgcgcc
301 ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag
361 ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgagacgaa agggcctcgt
421 gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg
481 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
541 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
601 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
661 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
721 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
781 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
841 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
901 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
961 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
1021 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
1081 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
1141 gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
1201 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
1261 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
1321 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
1381 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
1441 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
1501 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
1561 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
1621 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
1681 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
1741 gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta
1801 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
1861 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
1921 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
1981 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
2041 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
2101 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
2161 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
2221 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
2281 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
2341 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
2401 ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag
2461 ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag
2521 ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg
2581 tggaattgtg agcggataac aatttcacac aggaaacagc tatgaccatg attacgaatt
2641 tgggccgtct ctgcaaaagt ctaggcctcc aaaaaagcct cctcactact tctggaatag
2701 ctcagaggcc gaggcggcct cggcctctgc ataaataaaa aaaattagtc agccatgggg
2761 cggagaatgg gcggaactgg gcggagttag gggcgggata gctagagcca gacatgataa
2821 gatacattga tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt
2881 gtgaaatttg tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagttc
2941 ctctcactct ctgatattca tttctttgca agttataaat actgaataat aagatgacat
3001 gaactactac tgctagagat tttccacact gactaaaagg gtctgaggga tctctagtta
3061 ccagagtcac acaacagacg ggcacacact acttgaagca ctcaaggcaa gctttattga
3121 ggcttaagca gtgggttccc tagttagcca gagagctccc aggctcagat ctggtctaac
3181 cagagagacc cagtacaagc aaaaagcaga tcttattttc gttgggagtg aattagccct
3241 tccagtcccc ccttttcttt taaaaagtgg ctaagatcta cagctgcctt gtaagtcatt
3301 ggtcttaaag gtaccaggcg gggaggcggc ccaaagggag atccgactcg tctgagggcg
3361 aaggcgaaga cgcggaagag gccgcagagc cggcagcagg ccgcgggaag gaaggtccgc
3421 tggattgagg gccgaaggga cgtagcagaa ggacgtcccg cgcagaatcc aggtggcaac
3481 acaggcgagc agccaaggaa aggacgatga tttccccgac aacaccacgg aattgtcagt
3541 gcccaacagc cgagcccctg tccagcagcg ggcaaggcag gcggcgatga gttccgccgt
3601 ggcaataggg agggggaaag cgaaagtccc ggaaaggagc tgacaggtgg tggcaatgcc
3661 ccaaccagtg ggggttgcgt cagcaaacac agtgcacacc acgccacgtt gcctgacaac
3721 gggccacaac tcctcataaa gagacagcaa ccaggattta tacaaggagg agaaaatgaa
3781 agccatacgg gaagcaatag catgatacaa aggcattaaa gcagcgtatc cacatagcgt
3841 aaaaggagca acatagttaa gaataccagt caatctttca caaattttgt aatccagagg
3901 ttgattgtcg acttacttgt acagctcgtc catgccgtac aggaacaggt ggtggcggcc
3961 ctcggagcgc tcgtactgtt ccacgatggt gtagtcctcg ttgtgggagg tgatgtccag
4021 cttggtgtcc acgtagtagt agccgggcag ttgcacgggc ttcttggcca tgtagatggt
4081 cttgaactcc accaggtagt ggccgccgtc cttcagcttc agggcctggt ggatctcgcc
4141 cttcagcacg ccgtcgcggg ggtacaggcg ctcggtggag gcctcccagc ccatggtctt
4201 cttctgcatt acggggccgt cgggggggaa gttggtgccg cgcatcttca ccttgtagat
4261 cagcgtgccg tcctgcaggg aggagtcctg ggtcacggtc accagaccgc cgtcctcgaa
4321 gttcatcacg cgctcccact tgaagccctc ggggaaggac agcttcttgt aatcggggat
4381 gtcggcgggg tgcttcacgt acgccttgga gccgtacatg aactgggggg acaggatgtc
4441 ccaggcgaag ggcagggggc cgcccttggt caccttcagc ttggcggtct gggtgccctc
4501 gtaggggcgg ccctcgccct cgccctcgat ctcgaactcg tggccgttca tggagccctc
4561 catgcgcacc ttgaagcgca tgaactcttt gatgacggcc atgttgttgt cctcggagga
4621 ggcggtgccg gagctgccgc taccggtgct gccggtgcca tgccccagga acaggtggtg
4681 gcggccctcg gagcgctcgt actgttccac gatggtgtag tcctcgttgt gggaggtgat
4741 gtccagcttg gtgtccacgt agtagtagcc gggcagttgc acgggcttct tggccatgta
4801 gatggtcttg aactccacca ggtagtggcc gccgtccttc agcttcaggg cctggtggat
4861 ctcgcccttc agcacgccgt cgcgggggta caggcgctcg gtggaggcct cccagcccat
4921 ggtcttcttc tgcattacgg ggccgtcggg ggggaagttg gtgccgcgca tcttcacctt
4981 gtagatcagc gtgccgtcct gcagggagga gtcctgggtc acggtcacca gaccgccgtc
5041 ctcgaagttc atcacgcgct cccacttgaa gccctcgggg aaggacagct tcttgtaatc
5101 ggggatgtcg gcggggtgct tcacgtacgc cttggagccg tacatgaact ggggggacag
5161 gatgtcccag gcgaagggca gggggccgcc cttggtcacc ttcagcttgg cggtctgggt
5221 gccctcgtag gggcggccct cgccctcgcc ctcgatctcg aactcgtggc cgttcatgga
5281 gccctccatg cgcaccttga agcgcatgaa ctctttgatg acctcctcgc ccttgctcac
5341 catggttccg gaagggccgg gattctcctc cacgtcaccg catgttagaa gacttcctct
5401 gccctcagcg gccgctaaca cggcgatctt gccgcccttc ttggccttaa tgagaatctc
5461 gcggatcttg cgggcgtcca acttgccggt cagtccttta ggcacctcgt ccacgaacac
5521 aacaccaccg cgcagcttct tggcggttgt aacctggctg gccacatagt ccacgatctc
5581 cttctcggtc atggttttac cgtgttccag cacgacgact gcggcgggca gctcgccggc
5641 atcgtcgtcg ggcaggccgg cgaccccggc gtcgaagatg ttggggtgtt gcagcaggat
5701 gctctccagt tcggctgggg ctacctggta gcccttgtat ttgatcaggc tcttcagccg
5761 gtccacgatg aagaagtgct cgtcctcgtc ccagtaggcg atgtcgccgc tgtgcagcca
5821 gccgtccttg tcgatgagag cgtttgtagc ctcggggttg ttaacgtagc cgctcatgat
5881 catggggcca cggacgcaca gctcgccgcg ctggttcaca cccagtgtct taccggtgtc
5941 caagtccacc accttagcct cgaagaaggg caccaccttg cctactgcgc caggcttgtc
6001 gtccccttcg ggggtgatca gaatggcgct ggttgtttct gtcaggccgt agccctggcg
6061 gatgcctggt aggtggaagc gtttggccac ggcctcacct acctccttgc tgagcggcgc
6121 cccgccgctg gcgatctcgt gcaagttgct taggtcgtac ttgtcgatga gagtgctctt
6181 agcgaagaag ctaaatagtg tgggcaccag cagggcagat tgaatcttat agtcttgcaa
6241 gctgcgcaag aatagctcct cctcgaagcg gtacatgagc acgacccgaa agccgcagat
6301 caagtagccc agcgtggtga acatgccgaa gccgtggtga aatggcacca cgctgaggat
6361 agcggtgtcg gggatgatct ggttgccgaa gatggggtcg cgggcatgac tgaatcggac
6421 acaagcggtg cggtgcggta gggctacgcc cttgggcaat ccggtactgc cactactgtt
6481 catgatcagg gcgatggttt tgtcccggtc gaagctctcg ggcacgaagt cgtactcgtt
6541 gaagccgggt ggcaaatggg aagtcacgaa ggtgtacatg ctttggaagc cctggtagtc
6601 ggtcttgcta tccatgatga tgatcttttg tatgatcggt agcttctttt gcacgttgag
6661 gatcttttgc agccctttct tgctcacgaa tacgacggtg ggctggctga tgcccatgct
6721 gttcagcagc tcgcgctcgt tgtagatgtc gttagctggg gccacagcca caccgatgaa
6781 cagggcaccc aacacgggca tgaagaactg caagctattc tcgctgcaca ccacgatccg
6841 atggtttgta ttcagcccat agcgcttcat agcttctgcc agccgaacgc tcatctcgaa
6901 gtactcggcg taggtaatgt ccacctcgat atgtgcgtcg gtaaaggcga tggtgccggg
6961 caccagggcg tagcgcttca tggctttgtg cagctgctcg ccggcggtcc cgtcttcgag
7021 tgggtagaat ggcgctgggc ccttcttaat gtttttggca tcttccatga attcgctagc
7081 tctagagtag gcgccggtca cagcttggat ctgtaacggc gcagaacaga aaacgaaaca
7141 aagacgtaga gttgagcaag cagggtcagg caaagcgtgg agagccggct gagtctaggt
7201 aggctccaag ggagcgccgg acaaaggccc ggtctcgacc tgagctttaa acttacctag
7261 acggcggacg cagttcagga ggcaccacag gcgggaggcg gcagaacgcg actcaaccgg
7321 cgtggatggc ggcctcaggt agggcggcgg gcgcgtgaag gagagatgcg agcccctcga
7381 agcttcagct gtgttctggc ggcaaacccg ttgcgaaaaa gaacgttcac ggcgactact
7441 gcacttatat acggttctcc cccaccctcg ggaaaaaggc ggagccagta cacgacatca
7501 ctttcccagt ttaccccgcg ccaccttctc taggcacccg ttcaattgcc gacccctccc
7561 cccaacttct cggggactgt gggcgatgtg cgctctgccc actgacgggc accggagcga
7621 tcgcagatcc ttactagtat cgataaaatt ttgaattttt gtaatttgtt tttgtaattc
7681 tttagtttgt atgtctgttg ctattatgtc tactattctt tcccctgcac tgtacccccc
7741 aatcccccct tttcttttaa aagttaaccg ataccgtcga gatccgttca ctaatcgaat
7801 ggatctgtct ctgtctctct ctccaccttc ttcttctatt ccttcgggcc tgtcgggtcc
7861 cctcggggtt gggaggtggg tctgaaacga taatggtgaa tatccctgcc taactctatt
7921 cactatagaa agtacagcaa aaactattct taaacctacc aagcctccta ctatcattat
7981 gaataatttt atataccaca gccaatttgt tatgttaaac caattccaca aacttgccca
8041 tttatctaat tccaataatt cttgttcatt cttttcttgc tggttttgcg attcttcaat
8101 taaggagtgt attaagcttg tgtaattgtt aatttctctg tcccactcca tccaggtcgt
8161 gtgattccaa tctgttccag agatttatta ctccaactag cattccaagg cacagcagtg
8221 gtgcaaatga gttttccaga gcaaccccaa atccccagga gctgttgatc ctttaggtat
8281 ctttccacag ccaggattct tgcctggagc tgcttgatgc cccagactgt gagttgcaac
8341 agatgctgtt gcgcctcaat agccctcagc aaattgttct gctgctgcac tataccagac
8401 aataattgtc tggcctgtac cgtcagcgtc attgaggctg cgcccatagt gcttcctgct
8461 gctcccaaga acccaaggaa caaagctcct attcccactg ctcttttttc tctctgcacc
8521 actcttctct ttgccttggt gggtgctact cctaatggtt caatttttac tactttatat
8581 ttatataatt cacttctcca attgtccctc atatctcctc ctccaggtct gaagatcagt
8641 ggccgcttgc tgtgcggtgg tcttactttt gttttgctct tcctctatct tgtctaaagc
8701 ttccttggtg tcttttatct ctatcctttg atgcacacaa tagagggttg ctactgtatt
8761 atataatgat ctaagttctt ctgatcctgt ctgaagggat ggttgtagct gtcccagtat
8821 ttgtctacag ccttctgatg tttctaacag gccaggatta actgcgaatc gttctagctc
8881 cctgcttgcc catactatat gttttaattt atattttttc tttccccctg gccttaaccg
8941 aattttttcc catcgcgatc taattctccc ccgcttaata ctgacgctct cgcacccatc
9001 tctctccttc tagcctccgc tagtcaaaat ttttggcgta ctcaccagtc gccgcccctc
9061 gcctcttgcc gtgcgcgctt cagcaagccg agtcctgcgt cgagagagct ctggtttccc
9121 tttcgctttc aggtccctgt tcgggcgcca ctgctagaga ttttccacac tgactaaaag
9181 ggtctgaggg atctctagtt accagagtca cacaacagac gggcacacac tacttgaagc
9241 actcaaggca agctttattg aggcttaagc agtgggttcc ctagttagcc agagagctcc
9301 caggctcaga tctggtctaa ccagagagac ccgtttattg tatcgagcta ggcacttaaa
9361 tacaatatct ctgcaatgcg gcaattcagt ggttcgtcca atccatgtca gacccgtctg
9421 ttgccttcct aataaggcac gatcgtacca ccttacttcc accaatcggc atgcacggtg
9481 ctttttctct ccttgtaagg catgttgcta actcatcgtt accatgttgc aagactacaa
9541 gagtattgca taagactaca cgcgt
//