LOCUS V025964 6035 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V025964 VERSION V025964 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 941..1525 /label="IGF1(NM_001111285)" /note="IGF1(NM_001111285)" /gene="IGF1" CDS 1532..1597 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" polyA_signal 1635..1859 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 1905..2333 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 2347..2676 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 2743..3537 /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 3711..3832 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(3881..3897) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 3905..3921 /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(3929..3959) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 3974..3995 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(4283..4868) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5039..5899) /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(5900..6004) /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 atccgccacc atgggaaaaa tcagcagtct 961 tccaacccaa ttatttaagt gctgcttttg tgatttcttg aaggtgaaga tgcacaccat 1021 gtcctcctcg catctcttct acctggcgct gtgcctgctc accttcacca gctctgccac 1081 ggctggaccg gagacgctct gcggggctga gctggtggat gctcttcagt tcgtgtgtgg 1141 agacaggggc ttttatttca acaagcccac agggtatggc tccagcagtc ggagggcgcc 1201 tcagacaggc atcgtggatg agtgctgctt ccggagctgt gatctaagga ggctggagat 1261 gtattgcgca cccctcaagc ctgccaagtc agctcgctct gtccgtgccc agcgccacac 1321 cgacatgccc aagacccaga agtatcagcc cccatctacc aacaagaaca cgaagtctca 1381 gagaaggaaa ggttggccaa agacacatcc aggaggggaa cagaaggagg ggacagaagc 1441 aagtctgcag atcagaggaa agaagaaaga gcagaggagg gagattggaa gtagaaatgc 1501 tgaatgcaga ggcaaaaaag gaaaactcga ggactacaaa gaccatgacg gtgattataa 1561 agatcatgac atcgactaca aggatgacga tgacaagtag tgagggcccg tttaaacccg 1621 ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt 1681 gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat 1741 tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag 1801 caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc 1861 ttctgaggcg gaaagaacca gctggggctc tagggggtat ccccacgcgc cctgtagcgg 1921 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc 1981 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc 2041 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct 2101 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac 2161 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac 2221 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat 2281 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg 2341 tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg 2401 caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca 2461 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact 2521 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta 2581 atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag 2641 tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc 2701 attttcggat ctgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga 2761 ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa 2821 cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt 2881 ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg 2941 ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa 3001 gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac 3061 cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt 3121 gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact 3181 cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg 3241 ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg 3301 acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc 3361 atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt 3421 gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc 3481 gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg 3541 ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg 3601 attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct 3661 ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta 3721 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat 3781 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct 3841 gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt 3901 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag 3961 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt 4021 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag 4081 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg 4141 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat 4201 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta 4261 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa 4321 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc 4381 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt 4441 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca 4501 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg 4561 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat 4621 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta 4681 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct 4741 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac 4801 aaaccaccgc tggtagcggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag 4861 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact 4921 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa 4981 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt 5041 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag 5101 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca 5161 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc 5221 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt 5281 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg 5341 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca 5401 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg 5461 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca 5521 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg 5581 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct 5641 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca 5701 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca 5761 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg 5821 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac 5881 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt 5941 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc 6001 cgcgcacatt tccccgaaaa gtgccacctg acgtc //