LOCUS V040520 6745 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V040520 VERSION V040520 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 928..2181 /label="SSTR3(NM_001051)" /note="SSTR3(NM_001051)" /gene="SSTR3" CDS 2188..2211 /codon_start=1 /product="FLAG® epitope tag, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDDDDK" /label="FLAG" CDS 2218..2307 /codon_start=1 /product="two Strep-Tag®II moieties connected by a linker for enhanced binding to Strep-Tactin®, an engineered form of streptavidin (Schmidt et al., 2013)" /transl_table=1 /translation="SA,WSHPQFEK,GGGSGGGSGGSA,WSHPQFEK" /label="Twin-Strep-tag" polyA_signal 2345..2569 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 2615..3043 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 3057..3386 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3453..4247 /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 4421..4542 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4591..4607) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4615..4631 /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(4639..4669) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4684..4705 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(4993..5578) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5749..6609) /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(6610..6714) /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 aagctgaatt cgccaccatg gacatgcttc atccatcatc ggtgtccacg 961 acctcagaac ctgagaatgc ctcctcggcc tggcccccag atgccaccct gggcaacgtg 1021 tcggcgggcc caagcccggc agggctggcc gtcagtggcg ttctgatccc cctggtctac 1081 ctggtggtgt gcgtggtggg cctgctgggt aactcgctgg tcatctatgt ggtcctgcgg 1141 cacacggcca gcccttcagt caccaacgtc tacatcctca acctggcgct ggccgacgag 1201 ctcttcatgc tggggctgcc cttcctggcc gcccagaacg ccctgtccta ctggcccttc 1261 ggctccctca tgtgccgcct ggtcatggcg gtggatggca tcaaccagtt caccagcata 1321 ttctgcctga ctgtcatgag cgtggaccgc tacctggccg tggtacatcc cacccgctcg 1381 gcccgctggc gcacagctcc ggtggcccgc acggtcagcg cggctgtgtg ggtggcctca 1441 gccgtggtgg tgctgcccgt ggtggtcttc tcgggagtgc cccgcggcat gagcacctgc 1501 cacatgcagt ggcccgagcc ggcggcggcc tggcgagccg gcttcatcat ctacacggcc 1561 gcactgggct tcttcgggcc gctgctggtc atctgcctct gctacctgct catcgtggtg 1621 aaggtgcgct cagctgggcg ccgggtgtgg gcaccctcgt gccagcggcg gcggcgctcc 1681 gaacgcaggg tcacgcgcat ggtggtggcc gtggtggcgc tcttcgtgct ctgctggatg 1741 cccttctacg tgctcaacat cgtcaacgtg gtgtgcccac tgcccgagga gcctgccttc 1801 tttgggctct acttcctggt ggtggcgctg ccctatgcca acagctgtgc caaccccatc 1861 ctttatggct tcctctccta ccgcttcaag cagggcttcc gcagggtcct gctgcggccc 1921 tcccgccgtg tgcgcagcca ggagcccact gtggggcccc cggagaagac tgaggaggag 1981 gatgaggagg aggaggatgg ggaggagagc agggaggggg gcaaggggaa ggagatgaac 2041 ggccgggtca gccagatcac gcagcctggc accagcgggc aggagcggcc gcccagcaga 2101 gtggccagca aggagcagca gctcctaccc caagaggctt ccactgggga gaagtccagc 2161 acgatgcgca tcagctacct gctcgaggat tacaaggatg acgacgataa gggatccagc 2221 gcctggtccc accctcagtt tgagaagggc ggaggctctg gcggcggaag cggaggatct 2281 gcttggagcc acccccagtt cgaaaagtga tgagggcccg tttaaacccg ctgatcagcc 2341 tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg 2401 accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat 2461 tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag 2521 gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc ttctgaggcg 2581 gaaagaacca gctggggctc tagggggtat ccccacgcgc cctgtagcgg cgcattaagc 2641 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc 2701 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct 2761 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa 2821 aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc 2881 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca 2941 ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat 3001 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg tggaatgtgt 3061 gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc 3121 atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta 3181 tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc 3241 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta 3301 tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag tgaggaggct 3361 tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc attttcggat 3421 ctgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag 3481 gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg 3541 gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca 3601 agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc 3661 tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg 3721 actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg 3781 ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta 3841 cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag 3901 ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac 3961 tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg acccatggcg 4021 atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg 4081 gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg 4141 aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg 4201 attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg 4261 gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc 4321 cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct 4381 ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta ttgcagctta 4441 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact 4501 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtataccgtc 4561 gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta 4621 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc 4681 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg 4741 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg 4801 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg 4861 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa 4921 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc 4981 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc 5041 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag 5101 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct 5161 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta 5221 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc 5281 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc 5341 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt 5401 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct 5461 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc 5521 tggtagcggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga 5581 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg 5641 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg 5701 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt 5761 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact 5821 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat 5881 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg 5941 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg 6001 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat 6061 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc 6121 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt 6181 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc 6241 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga 6301 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc 6361 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa 6421 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta 6481 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg 6541 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg 6601 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat 6661 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt 6721 tccccgaaaa gtgccacctg acgtc //