LOCUS Exported 8189 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 8189)
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..8189
/organism="synthetic DNA construct"
/mol_type="other DNA"
LTR 234..414
/note="5' LTR (truncated)"
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
misc_feature 458..583
/note="HIV-1 Psi"
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 1076..1309
/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 1798..1915
/note="cPPT/CTS"
/note="central polypurine tract and central termination
sequence of HIV-1"
promoter 2005..2208
/note="CMV promoter"
/note="human cytomegalovirus (CMV) immediate early
promoter"
promoter 2348..2559
/note="EF-1-alpha core promoter"
/note="core promoter for human elongation factor
EF-1-alpha"
LTR 2572..2840
/note="5' LTR (truncated)"
/note="truncated 5' long terminal repeat (LTR) from human
T-cell leukemia virus (HTLV) type 1"
CDS 2899..3561
/codon_start=1
/product="green fluorescent protein 2 from Pontellina
plumata, also known as ppluGFP2 (Shagin et al., 2004)"
/note="CopGFP"
/translation="PAMEIECRITGTLNGVEFELVGGGEGTPKQGRMTNKMKSTKGALT
FSPYLLSHVMGYGFYHFGTYPSGYENPFLHAINNGGYTNTRIEKYEDGGVLHVSFSYRY
EAGRVIGDFKVVGTGFPEDSVIFTDKIIRSNATVEHLHPMGDNVLVGSFARTFSLRDGG
YYSFVVDSHMHFKSAIHPSILQNGGPMFAFRRVEELHSNTELGIVEYQHAFKTPIAFA"
CDS 3631..3684
/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 3685..4284
/codon_start=1
/gene="pac from Streptomyces alboniger"
/product="puromycin N-acetyltransferase"
/note="PuroR"
/note="confers resistance to puromycin"
/translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
misc_feature 4285..4872
/note="WPRE"
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
CDS complement(4756..4767)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/note="Factor Xa site"
/translation="IEGR"
LTR 4946..5179
/note="3' LTR (Delta-U3)"
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
polyA_signal 5251..5372
/note="SV40 poly(A) signal"
/note="SV40 polyadenylation signal"
rep_origin 5391..5526
/note="SV40 ori"
/note="SV40 origin of replication"
primer_bind complement(5564..5580)
/note="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 5588..5604
/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)."
promoter complement(5612..5642)
/note="lac promoter"
/note="promoter for the E. coli lac operon"
protein_bind 5657..5678
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(5966..6554)
/direction=LEFT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(6725..7585)
/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"
promoter complement(7586..7690)
/gene="bla"
/note="AmpR promoter"
primer_bind 8164..8180
/note="M13 fwd"
/note="common sequencing primer, one of multiple similar
variants"
ORIGIN
1 acgcgtgtag tcttatgcaa tactcttgta gtcttgcaac atggtaacga tgagttagca
61 acatgcctta caaggagaga aaaagcaccg tgcatgccga ttggtggaag taaggtggta
121 cgatcgtgcc ttattaggaa ggcaacagac gggtctgaca tggattggac gaaccactga
181 attgccgcat tgcagagata ttgtatttaa gtgcctagct cgatacaata aacgggtctc
241 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta
301 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact
361 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtggcg
421 cccgaacagg gacctgaaag cgaaagggaa accagagctc tctcgacgca ggactcggct
481 tgctgaagcg cgcacggcaa gaggcgaggg gcggcgactg gtgagtacgc caaaaatttt
541 gactagcgga ggctagaagg agagagatgg gtgcgagagc gtcagtatta agcgggggag
601 aattagatcg cgatgggaaa aaattcggtt aaggccaggg ggaaagaaaa aatataaatt
661 aaaacatata gtatgggcaa gcagggagct agaacgattc gcagttaatc ctggcctgtt
721 agaaacatca gaaggctgta gacaaatact gggacagcta caaccatccc ttcagacagg
781 atcagaagaa cttagatcat tatataatac agtagcaacc ctctattgtg tgcatcaaag
841 gatagagata aaagacacca aggaagcttt agacaagata gaggaagagc aaaacaaaag
901 taagaccacc gcacagcaag cggccactga tcttcagacc tggaggagga gatatgaggg
961 acaattggag aagtgaatta tataaatata aagtagtaaa aattgaacca ttaggagtag
1021 cacccaccaa ggcaaagaga agagtggtgc agagagaaaa aagagcagtg ggaataggag
1081 ctttgttcct tgggttcttg ggagcagcag gaagcactat gggcgcagcc tcaatgacgc
1141 tgacggtaca ggccagacaa ttattgtctg gtatagtgca gcagcagaac aatttgctga
1201 gggctattga ggcgcaacag catctgttgc aactcacagt ctggggcatc aagcagctcc
1261 aggcaagaat cctggctgtg gaaagatacc taaaggatca acagctcctg gggatttggg
1321 gttgctctgg aaaactcatt tgcaccactg ctgtgccttg gaatgctagt tggagtaata
1381 aatctctgga acagattgga atcacacgac ctggatggag tgggacagag aaattaacaa
1441 ttacacaagc ttaatacact ccttaattga agaatcgcaa aaccagcaag aaaagaatga
1501 acaagaatta ttggaattag ataaatgggc aagtttgtgg aattggttta acataacaaa
1561 ttggctgtgg tatataaaat tattcataat gatagtagga ggcttggtag gtttaagaat
1621 agtttttgct gtactttcta tagtgaatag agttaggcag ggatattcac cattatcgtt
1681 tcagacccac ctcccaaccc cgaggggacc cgacaggccc gaaggaatag aagaagaagg
1741 tggagagaga gacagagaca gatccattcg attagtgaac ggatctcgac ggttaacttt
1801 taaaagaaaa ggggggattg gggggtacag tgcaggggaa agaatagtag acataatagc
1861 aacagacata caaactaaag aattacaaaa acaaattaca aaaattcaaa attttatcga
1921 tactagtatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
1981 gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga
2041 tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg
2101 ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg
2161 caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctcg tttagtgaac
2221 cgtcagatcg cctggagacg ccatccacgc tgttttgacc tccatagaag attctagagc
2281 tagcgaattc gaatttaaat cggatccgcg gccgcaagga tctgcgatcg ctccggtgcc
2341 cgtcagtggg cagagcgcac atcgcccaca gtccccgaga agttgggggg aggggtcggc
2401 aattgaacgg gtgcctagag aaggtggcgc ggggtaaact gggaaagtga tgtcgtgtac
2461 tggctccgcc tttttcccga gggtggggga gaaccgtata taagtgcagt agtcgccgtg
2521 aacgttcttt ttcgcaacgg gtttgccgcc agaacacagc tgaagcttcg aggggctcgc
2581 atctctcctt cacgcgcccg ccgccctacc tgaggccgcc atccacgccg gttgagtcgc
2641 gttctgccgc ctcccgcctg tggtgcctcc tgaactgcgt ccgccgtcta ggtaagttta
2701 aagctcaggt cgagaccggg cctttgtccg gcgctccctt ggagcctacc tagactcagc
2761 cggctctcca cgctttgcct gaccctgctt gctcaactct acgtctttgt ttcgttttct
2821 gttctgcgcc gttacagatc caagctgtga ccggcgccta cgctagacgc caccatggag
2881 agcgacgaga gcggcctgcc cgccatggag atcgagtgcc gcatcaccgg caccctgaac
2941 ggcgtggagt tcgagctggt gggcggcgga gagggcaccc ccaagcaggg ccgcatgacc
3001 aacaagatga agagcaccaa aggcgccctg accttcagcc cctacctgct gagccacgtg
3061 atgggctacg gcttctacca cttcggcacc taccccagcg gctacgagaa ccccttcctg
3121 cacgccatca acaacggcgg ctacaccaac acccgcatcg agaagtacga ggacggcggc
3181 gtgctgcacg tgagcttcag ctaccgctac gaggccggcc gcgtgatcgg cgacttcaag
3241 gtggtgggca ccggcttccc cgaggacagc gtgatcttca ccgacaagat catccgcagc
3301 aacgccaccg tggagcacct gcaccccatg ggcgataacg tgctggtggg cagcttcgcc
3361 cgcaccttca gcctgcgcga cggcggctac tacagcttcg tggtggacag ccacatgcac
3421 ttcaagagcg ccatccaccc cagcatcctg cagaacgggg gccccatgtt cgccttccgc
3481 cgcgtggagg agctgcacag caacaccgag ctgggcatcg tggagtacca gcacgccttc
3541 aagaccccca tcgccttcgc cagatcccgc gctcagtcgt ccaattctgc cgtggacggc
3601 accgccggac ccggctccac cggatctcgc gagggcagag gaagtcttct aacatgcggt
3661 gacgtggagg agaatcccgg ccctatgacc gagtacaagc ccacggtgcg cctcgccacc
3721 cgcgacgacg tccccagggc cgtacgcacc ctcgccgccg cgttcgccga ctaccccgcc
3781 acgcgccaca ccgtcgatcc ggaccgccac atcgagcggg tcaccgagct gcaagaactc
3841 ttcctcacgc gcgtcgggct cgacatcggc aaggtgtggg tcgcggacga cggcgccgcg
3901 gtggcggtct ggaccacgcc ggagagcgtc gaagcggggg cggtgttcgc cgagatcggc
3961 ccgcgcatgg ccgagttgag cggttcccgg ctggccgcgc agcaacagat ggaaggcctc
4021 ctggcgccgc accggcccaa ggagcccgcg tggttcctgg ccaccgtcgg cgtctcgccc
4081 gaccaccagg gcaagggtct gggcagcgcc gtcgtgctcc ccggagtgga ggcggccgag
4141 cgcgccgggg tgcccgcctt cctggagacc tccgcgcccc gcaacctccc cttctacgag
4201 cggctcggct tcaccgtcac cgccgacgtc gaggtgcccg aaggaccgcg cacctggtgc
4261 atgacccgca agcccggtgc ctgaaatcaa cctctggatt acaaaatttg tgaaagattg
4321 actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct
4381 ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg
4441 ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact
4501 gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc
4561 gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc
4621 cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaag
4681 ctgacgtcct ttccatggct gctcgcctgt gttgccacct ggattctgcg cgggacgtcc
4741 ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg
4801 gctctgcggc ctcttccgcg tctccgcctt cgccctcaga cgagtcggat ctccctttgg
4861 ccgcctcccc gcctggtacc tttaagacca atgacttaca aggcagctgt agatcttagc
4921 cactttttaa aagaaaaggg gggactggaa gggctaattc actcccaacg aaaataagat
4981 ctgctttttg cttgtactgg gtctctctgg ttagaccaga tctgagcctg ggagctctct
5041 ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt gcttcaagta
5101 gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc cttttagtca
5161 gtgtggaaaa tctctagcag tagtagttca tgtcatctta ttattcagta tttataactt
5221 gcaaagaaat gaatatcaga gagtgagagg aacttgttta ttgcagctta taatggttac
5281 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
5341 tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggctctagct atcccgcccc
5401 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
5461 cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg
5521 gaggcctaga cttttgcaga gacggcccaa attcgtaatc atggtcatag ctgtttcctg
5581 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
5641 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
5701 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
5761 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
5821 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
5881 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
5941 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
6001 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
6061 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
6121 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
6181 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
6241 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
6301 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
6361 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta
6421 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
6481 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
6541 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
6601 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
6661 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
6721 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
6781 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
6841 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
6901 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
6961 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
7021 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
7081 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
7141 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
7201 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
7261 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
7321 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
7381 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
7441 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
7501 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
7561 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
7621 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
7681 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca
7741 tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg
7801 atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag
7861 cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg
7921 gctggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat atgcggtgtg
7981 aaataccgca cagatgcgta aggagaaaat accgcatcag gcgccattcg ccattcaggc
8041 tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctggcga
8101 aagggggatg tgctgcaagg cgattaagtt gggtaacgcc agggttttcc cagtcacgac
8161 gttgtaaaac gacggccagt gccaagctg
//