LOCUS Exported 8593 bp ds-DNA circular SYN 28-DEC-2017
DEFINITION synthetic circular DNA
FEATURES Location/Qualifiers
source 1..8593
/organism="synthetic DNA construct"
/mol_type="other DNA"
LTR 1..634
/label=3' LTR
/note="3' long terminal repeat (LTR) from HIV-1"
misc_feature 681..806
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 1303..1536
/label=RRE
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
misc_feature 2028..2143
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1 (lacking the first T)"
promoter 2196..2436
/label=U6 promoter
/note="RNA polymerase III promoter for human U6 snRNA"
promoter 2510..3009
/label=PGK promoter
/note="mouse phosphoglycerate kinase 1 promoter"
CDS 3036..3752
/codon_start=1
/product="enhanced GFP"
/label=EGFP
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
CDS 3768..3827
/codon_start=1
/product="2A peptide from equine rhinitis A virus
polyprotein"
/label=E2A
/note="Eukaryotic ribosomes fail to insert a peptide bond
between the Gly and Pro residues, yielding separate
polypeptides."
/translation="QCTNYALLKLAGDVESNPGP"
CDS 3828..4427
/codon_start=1
/gene="pac from Streptomyces alboniger"
/product="puromycin N-acetyltransferase"
/label=PuroR
/note="confers resistance to puromycin"
/translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
misc_feature 4521..5106
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
LTR 5310..5943
/label=3' LTR
/note="3' long terminal repeat (LTR) from HIV-1"
rep_origin complement(6473..7061)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(7232..8092)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
promoter complement(8093..8197)
/gene="bla"
/label=AmpR promoter
polyA_signal 8245..8379
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
ORIGIN
1 tggaagggct aattcactcc caaagaagac aagatatcct tgatctgtgg atctaccaca
61 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac
121 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca
181 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg
241 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag
301 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg
361 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat
421 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga
481 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
541 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
601 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag
661 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg
721 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga
781 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg
841 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg
901 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct
961 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat
1021 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca
1081 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc
1141 aagcggccgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag
1201 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc
1261 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg
1321 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc
1381 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc
1441 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct
1501 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa
1561 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca
1621 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt
1681 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt
1741 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta
1801 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt
1861 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct
1921 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga
1981 cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcgccttt aaaagaaaag
2041 gggggattgg ggggtacagt gcaggggaaa gaatagtaga cataatagca acagacatac
2101 aaactaaaga attacaaaaa caaattacaa aaattcaaaa ttttcgggtt tattacaggg
2161 acagcagaga tccagtttat cgatctgggc aggaagaggg cctatttccc atgattcctt
2221 catatttgca tatacgatac aaggctgtta gagagataat tagaattaat ttgactgtaa
2281 acacaaagat attagtacaa aatacgtgac gtagaaagta ataatttctt gggtagtttg
2341 cagttttaaa attatgtttt aaaatggact atcatatgct taccgtaact tgaaagtatt
2401 tcgatttctt ggctttatat atcttgtgga aaggacgagg atccgcctaa ggttaagtcg
2461 ccctcgctcg agcgagggcg acttaacctt aggttttttg aattctaccg ggtaggggag
2521 gcgcttttcc caaggcagtc tggagcatgc gctttagcag ccccgctggg cacttggcgc
2581 tacacaagtg gcctctggcc tcgcacacat tccacatcca ccggtaggcg ccaaccggct
2641 ccgttctttg gtggcccctt cgcgccacct tctactcctc ccctagtcag gaagttcccc
2701 cccgccccgc agctcgcgtc gtgcaggacg tgacaaatgg aagtagcacg tctcactagt
2761 ctcgtgcaga tggacagcac cgctgagcaa tggaagcggg taggcctttg gggcagcggc
2821 caatagcagc tttgctcctt cgctttctgg gctcagaggc tgggaagggg tgggtccggg
2881 ggcgggctca ggggcgggct caggggcggg gcgggcgccc gaaggtcctc cggaggcccg
2941 gcattctgca cgcttcaaaa gcgcacgtct gccgcgctgt tctcctcttc ctcatctccg
3001 ggcctttcga cctgcagccc aagcttaccg ccaccatggt gagcaagggc gaggagctgt
3061 tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca
3121 gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg aagttcatct
3181 gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctg acctacggcg
3241 tgcagtgctt cagccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca
3301 tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc aactacaaga
3361 cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca
3421 tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac tacaacagcc
3481 acaacgtcta tatcatggcc gacaagcaga agaacggcat caaggtgaac ttcaagatcc
3541 gccacaacat cgaggacggc agcgtgcagc tcgccgacca ctaccagcag aacaccccca
3601 tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagcacccag tccgccctga
3661 gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg accgccgccg
3721 ggatcactct cggcatggac gagctgtaca aggaatttgg cagcggccag tgcaccaact
3781 atgccctgct gaaactggct ggcgatgtgg aaagcaaccc cggacccatg accgagtaca
3841 agcccacggt gcgcctcgcc acccgcgacg acgtccccag ggccgtacgc accctcgccg
3901 ccgcgttcgc cgactacccc gccacgcgcc acaccgtcga tccggaccgc cacatcgagc
3961 gggtcaccga gctgcaagaa ctcttcctca cgcgcgtcgg gctcgacatc ggcaaggtgt
4021 gggtcgcgga cgacggcgcc gcggtggcgg tctggaccac gccggagagc gtcgaagcgg
4081 gggcggtgtt cgccgagatc ggcccgcgca tggccgagtt gagcggttcc cggctggccg
4141 cgcagcaaca gatggaaggc ctcctggcgc cgcaccggcc caaggagccc gcgtggttcc
4201 tggccaccgt cggcgtctcg cccgaccacc agggcaaggg tctgggcagc gccgtcgtgc
4261 tccccggagt ggaggcggcc gagcgcgccg gggtgcccgc cttcctggag acctccgcgc
4321 cccgcaacct ccccttctac gagcggctcg gcttcaccgt caccgccgac gtcgaggtgc
4381 ccgaaggacc gcgcacctgg tgcatgaccc gcaagcccgg tgcctgacgc ccgccccacg
4441 acccgcagcg cccgaccgaa aggagcgcac gaccccatgc atcggcgtct cgagtctaga
4501 gggcccgacg cgtctggaac aatcaacctc tggattacaa aatttgtgaa agattgactg
4561 gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta atggctttgt
4621 atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa tcctggttgc
4681 tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt
4741 ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc ctttccggga
4801 ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc cttgcccgct
4861 gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg gggaagctga
4921 cgtcctttcc atggctgctc gcctgtgttg ccacctggat tctgcgcggg acgtccttct
4981 gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg ctgccggctc
5041 tgcggcttcc gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct
5101 ccccgcctgg aattaattct gcagtcgaga cctagaaaaa catggagcaa tcacaagtag
5161 caatacagca gctatcaatg attgtgcctg gctagaagca caagaggagg aggaggtggg
5221 ttttccagtc acacctcagg tacctttaag accaatgact tacaaggcag ctgtagatct
5281 tagccacttt ttaaaagaaa agaggggact ggaagggcta attcactccc aacgaagaca
5341 agatatcctt gatctgtgga tctaccacac acaaggctac ttccctgatt agcagaacta
5401 cacaccaggg ccaggggtca gatatccact gacctttgga tggtgctaca agctagtacc
5461 agttgagcca gataaggtag aagaggccaa taaaggagag aacaccagct tgttacaccc
5521 tgtgagcctg catgggatgg atgacccgga gagagaagtg ttagagtgga ggtttgacag
5581 ccgcctagca tttcatcacg tggcccgaga gctgcatccg gagtacttca agaactgctg
5641 atatcgagct tgctacaagg gactttccgc tggggacttt ccagggaggc gtggcctggg
5701 cgggactggg gagtggcgag ccctcagatc ctgcatataa gcagctgctt tttgcctgta
5761 ctgggtctct ctggttagac cagatctgag cctgggagct ctctggctaa ctagggaacc
5821 cactgcttaa gcctcaataa agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt
5881 tgtgtgactc tggtaactag agatccctca gaccctttta gtcagtgtgg aaaatctcta
5941 gcagtagtag ttcatgtcat cttattattc agtatttata acttgcaaag aaatgaatat
6001 cagagagtga gaggccttga cattgctagc gtttaccgtc gacctctagc tagagcttgg
6061 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
6121 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
6181 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
6241 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
6301 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
6361 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
6421 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
6481 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
6541 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
6601 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
6661 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
6721 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
6781 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
6841 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
6901 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
6961 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
7021 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
7081 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
7141 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
7201 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
7261 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
7321 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
7381 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
7441 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
7501 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
7561 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
7621 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
7681 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
7741 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
7801 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
7861 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
7921 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
7981 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
8041 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
8101 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
8161 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
8221 ctgacgtcga cggatcggga gatcaacttg tttattgcag cttataatgg ttacaaataa
8281 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
8341 ttgtccaaac tcatcaatgt atcttatcat gtctggatca actggataac tcaagctaac
8401 caaaatcatc ccaaacttcc caccccatac cctattacca ctgccaatta cctgtggttt
8461 catttactct aaacctgtga ttcctctgaa ttattttcat tttaaagaaa ttgtatttgt
8521 taaatatgta ctacaaactt agtagttttt aaagaaattg tatttgttaa atatgtacta
8581 caaacttagt agt
//