LOCUS Exported 8215 bp ds-DNA circular SYN 01-DEC-2017
DEFINITION synthetic circular DNA
FEATURES Location/Qualifiers
source 1..8215
/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)"
enhancer 2201..2504
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 2505..2708
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
regulatory 2809..2818
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 2818..2883
/codon_start=1
/product="three tandem FLAG(R) epitope tags, followed by an
enterokinase cleavage site"
/label=3xFLAG
/translation="DYKDHDGDYKDHDIDYKDDDDK"
misc_feature 2884..2915
/label=MCS
misc_feature 2918..3491
/label=IRES
/note="internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)"
CDS 3524..4123
/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 4137..4725
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
LTR 4932..5565
/label=3' LTR
/note="3' long terminal repeat (LTR) from HIV-1"
rep_origin complement(6095..6683)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(6854..7714)
/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(7715..7819)
/gene="bla"
/label=AmpR promoter
polyA_signal 7867..8001
/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 cgataagctt gggagttccg cgttacataa cttacggtaa
2221 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg
2281 ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt
2341 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg
2401 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc
2461 ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc
2521 agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca
2581 ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta
2641 acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa
2701 gcagagctcg tttagtgaac cgtcagatcg cctggagacg ccatccacgc tgttttgacc
2761 tccatagaag acaccgactc tactagagga tctatttccg gtgaattcgc caccatggac
2821 tacaaagacc atgacggtga ttataaagat catgacatcg actacaagga tgacgatgac
2881 aagctcgaga ctagttctag agcggccgcg gatcccgccc ctctccctcc ccccccccta
2941 acgttactgg ccgaagccgc ttggaataag gccggtgtgc gtttgtctat atgttatttt
3001 ccaccatatt gccgtctttt ggcaatgtga gggcccggaa acctggccct gtcttcttga
3061 cgagcattcc taggggtctt tcccctctcg ccaaaggaat gcaaggtctg ttgaatgtcg
3121 tgaaggaagc agttcctctg gaagcttctt gaagacaaac aacgtctgta gcgacccttt
3181 gcaggcagcg gaacccccca cctggcgaca ggtgcctctg cggccaaaag ccacgtgtat
3241 aagatacacc tgcaaaggcg gcacaacccc agtgccacgt tgtgagttgg atagttgtgg
3301 aaagagtcaa atggctctcc tcaagcgtat tcaacaaggg gctgaaggat gcccagaagg
3361 taccccattg tatgggatct gatctggggc ctcggtgcac atgctttaca tgtgtttagt
3421 cgaggttaaa aaaacgtcta ggccccccga accacgggga cgtggttttc ctttgaaaaa
3481 cacgatgata agcttgccac aacccacaag gagacgacct tccatgaccg agtacaagcc
3541 cacggtgcgc ctcgccaccc gcgacgacgt cccccgggcc gtacgcaccc tcgccgccgc
3601 gttcgccgac taccccgcca cgcgccacac cgtcgacccg gaccgccaca tcgagcgggt
3661 caccgagctg caagaactct tcctcacgcg cgtcgggctc gacatcggca aggtgtgggt
3721 cgcggacgac ggcgccgcgg tggcggtctg gaccacgccg gagagcgtcg aagcgggggc
3781 ggtgttcgcc gagatcggcc cgcgcatggc cgagttgagc ggttcccggc tggccgcgca
3841 gcaacagatg gaaggcctcc tggcgccgca ccggcccaag gagcccgcgt ggttcctggc
3901 caccgtcggc gtctcgcccg accaccaggg caagggtctg ggcagcgccg tcgtgctccc
3961 cggagtggag gcggccgagc gcgccggggt gcccgccttc ctggagacct ccgcgccccg
4021 caacctcccc ttctacgagc ggctcggctt caccgtcacc gccgacgtcg aggtgcccga
4081 aggaccgcgc acctggtgca tgacccgcaa gcccggtgcc tagacgcgtc tggaacaatc
4141 aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt
4201 ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg
4261 ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc
4321 ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt
4381 ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg
4441 ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg
4501 gcactgacaa ttccgtggtg ttgtcgggga agctgacgtc ctttccatgg ctgctcgcct
4561 gtgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc
4621 cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc
4681 ttcgccctca gacgagtcgg atctcccttt gggccgcctc cccgcctgga attaattctg
4741 cagtcgagac ctagaaaaac atggagcaat cacaagtagc aatacagcag ctaccaatgc
4801 tgattgtgcc tggctagaag cacaagagga ggaggaggtg ggttttccag tcacacctca
4861 ggtaccttta agaccaatga cttacaaggc agctgtagat cttagccact ttttaaaaga
4921 aaagagggga ctggaagggc taattcactc ccaacgaaga caagatatcc ttgatctgtg
4981 gatctaccac acacaaggct acttccctga ttagcagaac tacacaccag ggccaggggt
5041 cagatatcca ctgacctttg gatggtgcta caagctagta ccagttgagc cagataaggt
5101 agaagaggcc aataaaggag agaacaccag cttgttacac cctgtgagcc tgcatgggat
5161 ggatgacccg gagagagaag tgttagagtg gaggtttgac agccgcctag catttcatca
5221 cgtggcccga gagctgcatc cggagtactt caagaactgc tgatatcgag cttgctacaa
5281 gggactttcc gctggggact ttccagggag gcgtggcctg ggcgggactg gggagtggcg
5341 agccctcaga tcctgcatat aagcagctgc tttttgcctg tactgggtct ctctggttag
5401 accagatctg agcctgggag ctctctggct aactagggaa cccactgctt aagcctcaat
5461 aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac tctggtaact
5521 agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtagt agttcatgtc
5581 atcttattat tcagtattta taacttgcaa agaaatgaat atcagagagt gagaggcctt
5641 gacattgcta gcgtttaccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc
5701 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
5761 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
5821 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
5881 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
5941 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
6001 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
6061 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
6121 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
6181 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
6241 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
6301 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
6361 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
6421 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
6481 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
6541 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
6601 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
6661 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
6721 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
6781 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
6841 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
6901 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
6961 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
7021 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
7081 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
7141 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
7201 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
7261 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
7321 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
7381 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
7441 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
7501 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
7561 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
7621 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
7681 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
7741 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
7801 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc gacggatcgg
7861 gagatcaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
7921 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
7981 gtatcttatc atgtctggat caactggata actcaagcta accaaaatca tcccaaactt
8041 cccaccccat accctattac cactgccaat tacctgtggt ttcatttact ctaaacctgt
8101 gattcctctg aattattttc attttaaaga aattgtattt gttaaatatg tactacaaac
8161 ttagtagttt ttaaagaaat tgtatttgtt aaatatgtac tacaaactta gtagt
//