LOCUS V039744 7507 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V039744 VERSION V039744 KEYWORDS . SOURCE . ORGANISM . . FEATURES Location/Qualifiers enhancer 235..614 /note="human cytomegalovirus immediate early enhancer" /label="CMV enhancer" promoter 615..818 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" promoter 863..881 /note="promoter for bacteriophage T7 RNA polymerase" /label="T7 promoter" CDS 964..2997 /label="GAS6(NM_000820)" /note="GAS6(NM_000820)" /gene="GAS6" CDS 3004..3069 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" polyA_signal 3107..3331 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 3377..3805 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 3819..4148 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 4215..5009 /codon_start=1 /gene="aph(3')-II (or nptII)" /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin®)" /product="aminoglycoside phosphotransferase from Tn5" /transl_table=1 /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*" /label="NeoR/KanR" polyA_signal 5183..5304 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(5353..5369) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 5377..5393 /bound_moiety="lac repressor encoded by lacI" /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-β-D-thiogalactopyranoside (IPTG)." /label="lac operator" promoter complement(5401..5431) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 5446..5467 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5755..6340) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(6511..7371) /codon_start=1 /gene="bla" /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /product="β-lactamase" /transl_table=1 /translation="MSIQHFRVALIPFFAAFCLPVFA,HPETLVKVKDAEDQLGARVGY IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEY SPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDR WEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRS ALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGA SLIKHW*" /label="AmpR" promoter complement(7372..7476) /gene="bla" /label="AmpR promoter" ORIGIN 1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg 61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg 121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc 181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt 241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata 301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc 361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc 421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt 481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt 541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca 601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg 661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc 721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg 781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca 841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc 901 gtttaaactt aagcttggta ccgagctcgg atccactagt ccagtgtggt ggaattcgcc 961 accatggccc cttcgctctc gcccgggccc gccgccctgc gccgcgcgcc gcagctgctg 1021 ctgctgctgc tggccgcgga gtgcgcgctt gccgcgctgt tgccggcgcg cgaggccacg 1081 cagttcctgc ggcccaggca gcgccgcgcc tttcaggtct tcgaggaggc caagcagggc 1141 cacctggaga gggagtgcgt ggaggagctg tgcagccgcg aggaggcgcg ggaggtgttc 1201 gagaacgacc ccgagacgga ttatttttac ccaagatact tagactgcat caacaagtat 1261 gggtctccgt acaccaaaaa ctcaggcttc gccacctgcg tgcaaaacct gcctgaccag 1321 tgcacgccca acccctgcga taggaagggg acccaagcct gccaggacct catgggcaac 1381 ttcttctgcc tgtgtaaagc tggctggggg ggccggctct gcgacaaaga tgtcaacgaa 1441 tgcagccagg agaacggggg ctgcctccag atctgccaca acaagccggg tagcttccac 1501 tgttcctgcc acagcggctt cgagctctcc tctgatggca ggacctgcca agacatagac 1561 gagtgcgcag actcggaggc ctgcggggag gcgcgctgca agaacctgcc cggctcctac 1621 tcctgcctct gtgacgaggg ctttgcgtac agctcccagg agaaggcttg ccgagatgtg 1681 gacgagtgtc tgcagggccg ctgtgagcag gtctgcgtga actccccagg gagctacacc 1741 tgccactgtg acgggcgtgg gggcctcaag ctgtcccagg acatggacac ctgtgaggac 1801 atcttgccgt gcgtgccctt cagcgtggcc aagagtgtga agtccttgta cctgggccgg 1861 atgttcagtg ggacccccgt gatccgactg cgcttcaaga ggctgcagcc caccaggctg 1921 gtagctgagt ttgacttccg gacctttgac cccgagggca tcctcctctt tgccggaggc 1981 caccaggaca gcacctggat cgtgctggcc ctgagagccg gccggctgga gctgcagctg 2041 cgctacaacg gtgtcggccg tgtcaccagc agcggcccgg tcatcaacca tggcatgtgg 2101 cagacaatct ctgttgagga gctggcgcgg aatctggtca tcaaggtcaa cagggatgct 2161 gtcatgaaaa tcgcggtggc cggggacttg ttccaaccgg agcgaggact gtatcatctg 2221 aacctgaccg tgggaggtat tcccttccat gagaaggacc tcgtgcagcc tataaaccct 2281 cgtctggatg gctgcatgag gagctggaac tggctgaacg gagaagacac caccatccag 2341 gaaacggtga aagtgaacac gaggatgcag tgcttctcgg tgacggagag aggctctttc 2401 taccccggga gcggcttcgc cttctacagc ctggactaca tgcggacccc tctggacgtc 2461 gggactgaat caacctggga agtagaagtc gtggctcaca tccgcccagc cgcagacaca 2521 ggcgtgctgt ttgcgctctg ggcccccgac ctccgtgccg tgcctctctc tgtggcactg 2581 gtagactatc actccacgaa gaaactcaag aagcagctgg tggtcctggc cgtggagcat 2641 acggccttgg ccctaatgga gatcaaggtc tgcgacggcc aagagcacgt ggtcaccgtc 2701 tcgctgaggg acggtgaggc caccctggag gtggacggca ccaggggcca gagcgaggtg 2761 agcgccgcgc agctgcagga gaggctggcc gtgctcgaga ggcacctgcg gagccccgtg 2821 ctcacctttg ctggcggcct gccagatgtg ccggtgactt cagcgccagt caccgcgttc 2881 taccgcggct gcatgacact ggaggtcaac cggaggctgc tggacctgga cgaggcggcg 2941 tacaagcaca gcgacatcac ggcccactcc tgcccccccg tggagcccgc cgcagccctc 3001 gaggactaca aagaccatga cggtgattat aaagatcatg acatcgacta caaggatgac 3061 gatgacaagt agtgagggcc cgtttaaacc cgctgatcag cctcgactgt gccttctagt 3121 tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact 3181 cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat 3241 tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc 3301 aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac cagctggggc 3361 tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt 3421 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc 3481 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct 3541 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat 3601 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc 3661 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc 3721 tattcttttg atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg 3781 atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa 3841 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa 3901 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca 3961 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca 4021 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg 4081 ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct 4141 tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcaa gagacaggat 4201 gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg 4261 tggagaggct attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg 4321 tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg 4381 ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc 4441 cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg 4501 aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca 4561 tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc 4621 aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg 4681 atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg 4741 cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata 4801 tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg 4861 accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat 4921 gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct 4981 tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca 5041 agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt 5101 gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat 5161 gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag 5221 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt 5281 gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt 5341 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca 5401 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact 5461 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct 5521 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc 5581 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca 5641 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg 5701 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca 5761 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa 5821 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc 5881 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc 5941 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct 6001 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg 6061 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag 6121 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta 6181 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg 6241 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtttttttgt 6301 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc 6361 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt 6421 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta 6481 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat 6541 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac 6601 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg 6661 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag 6721 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt 6781 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt 6841 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt 6901 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt 6961 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct 7021 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt 7081 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac 7141 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa 7201 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa 7261 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca 7321 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct 7381 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga 7441 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc 7501 tgacgtc //