LOCUS V032054 6008 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V032054 VERSION V032054 KEYWORDS . SOURCE . ORGANISM . . FEATURES Location/Qualifiers enhancer 202..581 /note="human cytomegalovirus immediate early enhancer" /label="CMV enhancer" promoter 582..785 /note="human cytomegalovirus (CMV) immediate early promoter" /label="CMV promoter" promoter 830..848 /note="promoter for bacteriophage T7 RNA polymerase" /label="T7 promoter" CDS 908..1501 /label="Sdc4(NM_011521)" /note="Sdc4(NM_011521)" /gene="Sdc4" CDS 1511..1528 /codon_start=1 /product="6xHis affinity tag" /transl_table=1 /translation="HHHHHH" /label="6xHis" polyA_signal 1575..1799 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 1845..2273 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 2287..2616 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 2683..3477 /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 3651..3772 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(3821..3837) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 3845..3861 /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(3869..3899) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 3914..3935 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(4223..4808) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(4979..5839) /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(5840..5944) /gene="bla" /label="AmpR promoter" ORIGIN 1 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt 61 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct 121 tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg 181 tacgggccag atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta 241 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg 301 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc 361 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa 421 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca 481 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta 541 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt 601 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg 661 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca 721 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca 781 gagctctctg gctaactaga gaacccactg cttactggct tatcgaaatt aatacgactc 841 actataggga gacccaagct ggctagcgtt taaacttaag cttggtaccg agctcggatc 901 cgccaccatg gcgcctgcct gcctgcttgc gccgctgctg ctgctgctcc tcggaggctt 961 ccccttggtc cccggagagt cgattcgaga gacagaggtc atcgaccccc aggacctcct 1021 ggaaggcaga tacttctctg gagccctccc cgacgacgaa gatgctggcg gctcggatga 1081 ctttgagctc tcgggttctg gagatctgga tgacacggag gagcccaggc ccttccctga 1141 agtgattgag cccttggtgc cactggataa ccacatccct gagaatgcac agcctggcat 1201 ccgtgtcccc tcagagccca aggaactgga agagaatgag gtcattccta aaagggcccc 1261 ctccgacgtg ggggatgaca tgtccaacaa agtatccatg tccagcactg cccagggcag 1321 caacatcttt gagagaactg aggtcttggc agctctgatc gtgggcggcg tggtaggcat 1381 cctctttgcc gttttcctga tcctgctgct ggtgtaccgc atgaagaaga aggacgaagg 1441 cagttacgac ttgggcaaga aacccatcta caaaaaagcc cccaccaatg agttctacgc 1501 agcggccgca catcaccatc atcaccattg actcgagtct agagggcccg tttaaacccg 1561 ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt 1621 gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat 1681 tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag 1741 caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc 1801 ttctgaggcg gaaagaacca gctggggctc tagggggtat ccccacgcgc cctgtagcgg 1861 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc 1921 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc 1981 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct 2041 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac 2101 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac 2161 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat 2221 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg 2281 tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg 2341 caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca 2401 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact 2461 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta 2521 atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag 2581 tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc 2641 attttcggat ctgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga 2701 ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa 2761 cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt 2821 ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg 2881 ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa 2941 gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac 3001 cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt 3061 gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact 3121 cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg 3181 ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg 3241 acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc 3301 atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt 3361 gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc 3421 gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg 3481 ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg 3541 attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct 3601 ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta 3661 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat 3721 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct 3781 gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt 3841 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag 3901 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt 3961 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag 4021 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg 4081 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat 4141 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta 4201 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa 4261 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc 4321 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt 4381 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca 4441 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg 4501 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat 4561 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta 4621 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct 4681 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac 4741 aaaccaccgc tggtagcggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag 4801 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact 4861 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa 4921 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt 4981 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag 5041 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca 5101 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc 5161 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt 5221 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg 5281 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca 5341 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg 5401 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca 5461 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg 5521 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct 5581 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca 5641 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca 5701 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg 5761 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac 5821 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt 5881 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc 5941 cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg atcgggagat ctcccgatcc 6001 cctatggt //