LOCUS V035021 5962 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V035021 VERSION V035021 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..1452 /label="Hint2(NM_026871)" /note="Hint2(NM_026871)" /gene="Hint2" CDS 1459..1524 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" polyA_signal 1562..1786 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 1832..2260 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 2274..2603 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 2670..3464 /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 3638..3759 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(3808..3824) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 3832..3848 /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(3856..3886) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 3901..3922 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(4210..4795) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(4966..5826) /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(5827..5931) /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 accatggctg ctgctgtgct gctggccgtg ggcttgcgcg ctgcgcgcag aaccctggct 1021 gccgcggggg cccggggggc gcaggtccga ggaaacgcag gggtgtccga cgggagtgaa 1081 gtggccaagg cccagaaagc agctcctgga ggagcatcgc caaccatctt ctcccggatt 1141 ttggaccgga gtctcccagc tgacattcta tatgaagatc agcagtgcct tgtgttccgt 1201 gacgtagctc ctcaggctcc tgtgcacttt ctggtcattc ctaggaagcc cattcctcga 1261 attagccagg ctgaagagga cgaccagcag cttctaggac acttactcct tgtggccaag 1321 aagatagcac aggcccaggg tctgaaagat gggtatcgac ttgtggttaa tgatggcaag 1381 atgggcgcac agtctgtgta tcacctgcac attcacgtac ttggaggccg acagctgcag 1441 tggccacctg gcctcgagga ctacaaagac catgacggtg attataaaga tcatgacatc 1501 gactacaagg atgacgatga caagtagtga gggcccgttt aaacccgctg atcagcctcg 1561 actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc 1621 ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt 1681 ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat 1741 tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa 1801 agaaccagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc attaagcgcg 1861 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct 1921 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta 1981 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa 2041 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct 2101 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc 2161 aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg 2221 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc 2281 agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc 2341 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc 2401 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc 2461 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt 2521 atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt 2581 ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt ttcggatctg 2641 atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt 2701 ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct 2761 gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga 2821 ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg 2881 ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact 2941 ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg 3001 agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct 3061 gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg 3121 gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt 3181 tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg 3241 cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc 3301 ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag 3361 agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt 3421 cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt 3481 cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc 3541 cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca 3601 gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa 3661 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca 3721 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac 3781 ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc 3841 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta 3901 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa 3961 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat 4021 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg 4081 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc 4141 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt 4201 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag 4261 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc 4321 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc 4381 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt 4441 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt 4501 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc 4561 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa 4621 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa 4681 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg 4741 tagcggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga 4801 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat 4861 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag 4921 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat 4981 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc 5041 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat 5101 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag 5161 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg 5221 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc 5281 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca 5341 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg 5401 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc 5461 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta 5521 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc 5581 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg 5641 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc 5701 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc 5761 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat 5821 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag 5881 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc 5941 ccgaaaagtg ccacctgacg tc //