LOCUS Exported 8500 bp ds-DNA circular SYN 06-SEP-2016
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS Untitled 13
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8500)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Tuesday, September 6, 2016 from SnapGene Viewer 3.1.4
http://www.miaolingbio.com
FEATURES Location/Qualifiers
source 1..8500
/organism="synthetic DNA construct"
/mol_type="other DNA"
misc_feature 580..921
/note="MESV Psi"
/note="packaging signal of murine embryonic stem cell
virus"
CDS 988..1404
/note="gag (truncated)"
/note="truncated Moloney murine leukemia virus (MMLV) gag
gene lacking the start codon"
protein_bind 1438..1471
/bound_moiety="Cre recombinase"
/note="loxP"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (GCATACAT)."
CDS 1506..2183
/codon_start=1
/product="noncytotoxic tetrameric variant of DsRed
fluorescent protein (Strack et al., 2008)"
/note="DsRed-Express2"
/note="mammalian codon-optimized"
/translation="MDSTENVIKPFMRFKVHMEGSVNGHEFEIEGEGEGKPYEGTQTAK
LQVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGV
VTVTQDSSLQDGTFIYHVKFIGVNFPSDGPVMQKKTLGWEPSTERLYPRDGVLKGEIHK
ALKLKGGGHYLVEFKSIYMAKKPVKLPGYYYVDSKLDITSHNEDYTVVEQYERAEARHH
LFQ"
protein_bind 2208..2241
/bound_moiety="Cre recombinase"
/note="loxP"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (GCATACAT)."
CDS 2298..3017
/codon_start=1
/product="enhanced GFP"
/note="EGFP"
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
promoter 3027..3526
/note="PGK promoter"
/note="mouse phosphoglycerate kinase 1 promoter"
CDS 3547..4146
/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 4216..4804
/note="WPRE"
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
CDS complement(4687..4698)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/note="Factor Xa site"
/translation="IEGR"
LTR 4861..5376
/note="3' LTR"
/note="3' long terminal repeat from murine embryonic stem
cell virus"
primer_bind complement(5545..5561)
/note="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 5569..5585
/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(5593..5623)
/note="lac promoter"
/note="promoter for the E. coli lac operon"
protein_bind 5638..5659
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(5947..6535)
/direction=LEFT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(6706..7566)
/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(7567..7671)
/gene="bla"
/note="AmpR promoter"
LTR 8499..516
/note="5' LTR"
/note="5' long terminal repeat from murine embryonic stem
cell virus"
ORIGIN
1 tgaaagaccc cacctgtagg tttggcaagc tagcttaagt aacgccattt tgcaaggcat
61 ggaaaataca taactgagaa tagagaagtt cagatcaagg ttaggaacag agagacagca
121 gaatatgggc caaacaggat atctgtggta agcagttcct gccccggctc agggccaaga
181 acagatggtc cccagatgcg gtcccgccct cagcagtttc tagagaacca tcagatgttt
241 ccagggtgcc ccaaggacct gaaaatgacc ctgtgcctta tttgaactaa ccaatcagtt
301 cgcttctcgc ttctgttcgc gcgcttctgc tccccgagct caataaaaga gcccacaacc
361 cctcactcgg cgcgccagtc ctccgataga ctgcgtcgcc cgggtacccg tattcccaat
421 aaagcctctt gctgtttgca tccgaatcgt ggactcgctg atccttggga gggtctcctc
481 agattgattg actgcccacc tcgggggtct ttcatttgga ggttccaccg agatttggag
541 acccctgcct agggaccacc gacccccccg ccgggaggta agctggccag cggtcgtttc
601 gtgtctgtct ctgtctttgt gcgtgtttgt gccggcatct aatgtttgcg cctgcgtctg
661 tactagttag ctaactagct ctgtatctgg cggacccgtg gtggaactga cgagttctga
721 acacccggcc gcaaccctgg gagacgtccc agggactttg ggggccgttt ttgtggcccg
781 acctgaggaa gggagtcgat gtggaatccg accccgtcag gatatgtggt tctggtagga
841 gacgagaacc taaaacagtt cccgcctccg tctgaatttt tgctttcggt ttggaaccga
901 agccgcgcgt cttgtctgct gcagcgctgc agcatcgttc tgtgttgtct ctgtctgact
961 gtgtttctgt atttgtctga aaattagggc cagactgtta ccactccctt aagtttgacc
1021 ttaggtcact ggaaagatgt cgagcggatc gctcacaacc agtcggtaga tgtcaagaag
1081 agacgttggg ttaccttctg ctctgcagaa tggccaacct ttaacgtcgg atggccgcga
1141 gacggcacct ttaaccgaga cctcatcacc caggttaaga tcaaggtctt ttcacctggc
1201 ccgcatggac acccagacca ggtcccctac atcgtgacct gggaagcctt ggcttttgac
1261 ccccctccct gggtcaagcc ctttgtacac cctaagcctc cgcctcctct tcctccatcc
1321 gccccgtctc tcccccttga acctcctcgt tcgaccccgc ctcgatcctc cctttatcca
1381 gccctcactc cttctctagg cgccggaatt agatcactag tccagtgtgg tggaattata
1441 acttcgtata gcatacatta tacgaagtta taattctgca gatatccagc acagtggcgg
1501 ccaccatgga tagcactgag aacgtcatca agcccttcat gcgcttcaag gtgcacatgg
1561 agggctccgt gaacggccac gagttcgaga tcgagggcga gggcgagggc aagccctacg
1621 agggcaccca gaccgccaag ctgcaggtga ccaagggcgg ccccctgccc ttcgcctggg
1681 acatcctgtc cccccagttc cagtacggct ccaaggtgta cgtgaagcac cccgccgaca
1741 tccccgacta caagaagctg tccttccccg agggcttcaa gtgggagcgc gtgatgaact
1801 tcgaggacgg cggcgtggtg accgtgaccc aggactcctc cctgcaggac ggcaccttca
1861 tctaccacgt gaagttcatc ggcgtgaact tcccctccga cggccccgta atgcagaaga
1921 agactctggg ctgggagccc tccaccgagc gcctgtaccc ccgcgacggc gtgctgaagg
1981 gcgagatcca caaggcgctg aagctgaagg gcggcggcca ctacctggtg gagttcaagt
2041 caatctacat ggccaagaag cccgtgaagc tgcccggcta ctactacgtg gactccaagc
2101 tggacatcac ctcccacaac gaggactaca ccgtggtgga gcagtacgag cgcgccgagg
2161 cccgccacca cctgttccag tagagctagc tgaataaggc cgctcgaata acttcgtata
2221 gcatacatta tacgaagtta ttcgagtcta gagggcccgt tagatctctc gaggttaacg
2281 aattcgacta cgccaccatg gtgagcaagg gcgaggagct gttcaccggg gtggtgccca
2341 tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg
2401 agggcgatgc cacctacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc
2461 ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct
2521 accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc
2581 aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt
2641 tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg
2701 gcaacatcct ggggcacaag ctggagtaca actacaacag ccacaacgtc tatatcatgg
2761 ccgacaagca gaagaacggc atcaaggtga acttcaagat ccgccacaac atcgaggacg
2821 gcagcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc
2881 tgctgcccga caaccactac ctgagcaccc agtccgccct gagcaaagac cccaacgaga
2941 agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg
3001 acgagctgta caagtaaaat tctaccgggt aggggaggcg cttttcccaa ggcagtctgg
3061 agcatgcgct ttagcagccc cgctgggcac ttggcgctac acaagtggcc tctggcctcg
3121 cacacattcc acatccaccg gtaggcgcca accggctccg ttctttggtg gccccttcgc
3181 gccaccttct actcctcccc tagtcaggaa gttccccccc gccccgcagc tcgcgtcgtg
3241 caggacgtga caaatggaag tagcacgtct cactagtctc gtgcagatgg acagcaccgc
3301 tgagcaatgg aagcgggtag gcctttgggg cagcggccaa tagcagcttt gctccttcgc
3361 tttctgggct cagaggctgg gaaggggtgg gtccgggggc gggctcaggg gcgggctcag
3421 gggcggggcg ggcgcccgaa ggtcctccgg aggcccggca ttctgcacgc ttcaaaagcg
3481 cacgtctgcc gcgctgttct cctcttcctc atctccgggc ctttcgacct gcagcccaag
3541 cttaccatga ccgagtacaa gcccacggtg cgcctcgcca cccgcgacga cgtccccagg
3601 gccgtacgca ccctcgccgc cgcgttcgcc gactaccccg ccacgcgcca caccgtcgat
3661 ccggaccgcc acatcgagcg ggtcaccgag ctgcaagaac tcttcctcac gcgcgtcggg
3721 ctcgacatcg gcaaggtgtg ggtcgcggac gacggcgccg cggtggcggt ctggaccacg
3781 ccggagagcg tcgaagcggg ggcggtgttc gccgagatcg gcccgcgcat ggccgagttg
3841 agcggttccc ggctggccgc gcagcaacag atggaaggcc tcctggcgcc gcaccggccc
3901 aaggagcccg cgtggttcct ggccaccgtc ggcgtctcgc ccgaccacca gggcaagggt
3961 ctgggcagcg ccgtcgtgct ccccggagtg gaggcggccg agcgcgccgg ggtgcccgcc
4021 ttcctggaga cctccgcgcc ccgcaacctc cccttctacg agcggctcgg cttcaccgtc
4081 accgccgacg tcgaggtgcc cgaaggaccg cgcacctggt gcatgacccg caagcccggt
4141 gcctgacgcc cgccccacga cccgcagcgc ccgaccgaaa ggagcgcacg accccatgca
4201 tcgataattc ccgataatca acctctggat tacaaaattt gtgaaagatt gactggtatt
4261 cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat
4321 gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct
4381 ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct
4441 gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc
4501 gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg
4561 acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa gctgacgtcc
4621 tttccatggc tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac
4681 gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg
4741 cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc
4801 ccgcatcggg aattatcgat aaaataaaag attttattta gtctccagaa aaagggggga
4861 atgaaagacc ccacctgtag gtttggcaag ctagcttaag taacgccatt ttgcaaggca
4921 tggaaaatac ataactgaga atagagaagt tcagatcaag gttaggaaca gagagacagc
4981 agaatatggg ccaaacagga tatctgtggt aagcagttcc tgccccggct cagggccaag
5041 aacagatggt ccccagatgc ggtcccgccc tcagcagttt ctagagaacc atcagatgtt
5101 tccagggtgc cccaaggacc tgaaaatgac cctgtgcctt atttgaacta accaatcagt
5161 tcgcttctcg cttctgttcg cgcgcttctg ctccccgagc tcaataaaag agcccacaac
5221 ccctcactcg gcgcgccagt cctccgatag actgcgtcgc ccgggtaccc gtgtatccaa
5281 taaaccctct tgcagttgca tccgacttgt ggtctcgctg ttccttggga gggtctcctc
5341 tgagtgattg actacccgtc agcgggggtc tttcatgggt aacagtttct tgaagttgga
5401 gaacaacatt ctgagggtag gagtcgaata ttaagtaatc ctgactcaat tagccactgt
5461 tttgaatcca catactccaa tactcctgaa atagttcatt atggacagcg cagaagagct
5521 ggggagaatt aattcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
5581 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
5641 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
5701 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
5761 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
5821 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
5881 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
5941 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
6001 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
6061 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
6121 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
6181 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
6241 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
6301 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
6361 gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc
6421 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
6481 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
6541 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
6601 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
6661 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
6721 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
6781 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
6841 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
6901 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
6961 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
7021 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
7081 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
7141 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
7201 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
7261 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
7321 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
7381 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
7441 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
7501 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
7561 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
7621 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
7681 ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa
7741 taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg
7801 acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca
7861 agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta actatgcggc
7921 atcagagcag attgtactga gagtgcacca tatgcggtgt gaaataccgc acagatgcgt
7981 aaggagaaaa taccgcatca ggcgccattc gccattcagg ctgcgcaact gttgggaagg
8041 gcgatcggtg cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag
8101 gcgattaagt tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggcgca
8161 aggaatggtg catgcaagga gatggcgccc aacagtcccc cggccacggg gcctgccacc
8221 atacccacgc cgaaacaagc gctcatgagc ccgaagtggc gagcccgatc ttccccatcg
8281 gtgatgtcgg cgatataggc gccagcaacc gcacctgtgg cgccggtgat gccggccacg
8341 atgcgtccgg cgtagaggcg attagtccaa tttgttaaag acaggatatc agtggtccag
8401 gctctagttt tgactcaaca atatcaccag ctgaagccta tagagtacga gccatagata
8461 aaataaaaga ttttatttag tctccagaaa aaggggggaa
//