LOCUS V037518 6862 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V037518 VERSION V037518 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..2352 /label="GLP1R(NM_002062)" /note="GLP1R(NM_002062)" /gene="GLP1R" CDS 2359..2424 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" polyA_signal 2462..2686 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 2732..3160 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 3174..3503 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3570..4364 /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 4538..4659 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4708..4724) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4732..4748 /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(4756..4786) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4801..4822 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5110..5695) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5866..6726) /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(6727..6831) /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 accatggccg gcgcccccgg cccgctgcgc cttgcgctgc tgctgctcgg gatggtgggc 1021 agggccggcc cccgccccca gggtgccact gtgtccctct gggagacggt gcagaaatgg 1081 cgagaatacc gacgccagtg ccagcgctcc ctgactgagg atccacctcc tgccacagac 1141 ttgttctgca accggacctt cgatgaatac gcctgctggc cagatgggga gccaggctcg 1201 ttcgtgaatg tcagctgccc ctggtacctg ccctgggcca gcagtgtgcc gcagggccac 1261 gtgtaccggt tctgcacagc tgaaggcctc tggctgcaga aggacaactc cagcctgccc 1321 tggagggact tgtcggagtg cgaggagtcc aagcgagggg aaagaagctc cccggaggag 1381 cagctcctgt tcctctacat catctacacg gtgggctacg cactctcctt ctctgctctg 1441 gttatcgcct ctgcgatcct cctcggcttc agacacctgc actgcaccag gaactacatc 1501 cacctgaacc tgtttgcatc cttcatcctg cgagcattgt ccgtcttcat caaggacgca 1561 gccctgaagt ggatgtatag cacagccgcc cagcagcacc agtgggatgg gctcctctcc 1621 taccaggact ctctgagctg ccgcctggtg tttctgctca tgcagtactg tgtggcggcc 1681 aattactact ggctcttggt ggagggcgtg tacctgtaca cactgctggc cttctcggtc 1741 ttatctgagc aatggatctt caggctctac gtgagcatag gctggggtgt tcccctgctg 1801 tttgttgtcc cctggggcat tgtcaagtac ctctatgagg acgagggctg ctggaccagg 1861 aactccaaca tgaactactg gctcattatc cggctgccca ttctctttgc cattggggtg 1921 aacttcctca tctttgttcg ggtcatctgc atcgtggtat ccaaactgaa ggccaatctc 1981 atgtgcaaga cagacatcaa atgcagactt gccaagtcca cgctgacact catccccctg 2041 ctggggactc atgaggtcat ctttgccttt gtgatggacg agcacgcccg ggggaccctg 2101 cgcttcatca agctgtttac agagctctcc ttcacctcct tccaggggct gatggtggcc 2161 atattatact gctttgtcaa caatgaggtc cagctggaat ttcggaagag ctgggagcgc 2221 tggcggcttg agcacttgca catccagagg gacagcagca tgaagcccct caagtgtccc 2281 accagcagcc tgagcagtgg agccacggcg ggcagcagca tgtacacagc cacttgccag 2341 gcctcctgca gcctcgagga ctacaaagac catgacggtg attataaaga tcatgacatc 2401 gactacaagg atgacgatga caagtagtga gggcccgttt aaacccgctg atcagcctcg 2461 actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc 2521 ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt 2581 ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat 2641 tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa 2701 agaaccagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc attaagcgcg 2761 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct 2821 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta 2881 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa 2941 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct 3001 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc 3061 aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg 3121 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc 3181 agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc 3241 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc 3301 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc 3361 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt 3421 atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt 3481 ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt ttcggatctg 3541 atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt 3601 ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct 3661 gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga 3721 ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg 3781 ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact 3841 ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg 3901 agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct 3961 gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg 4021 gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt 4081 tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg 4141 cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc 4201 ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag 4261 agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt 4321 cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt 4381 cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc 4441 cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca 4501 gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa 4561 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca 4621 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac 4681 ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc 4741 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta 4801 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa 4861 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat 4921 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg 4981 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc 5041 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt 5101 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag 5161 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc 5221 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc 5281 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt 5341 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt 5401 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc 5461 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa 5521 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa 5581 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg 5641 tagcggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga 5701 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat 5761 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag 5821 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat 5881 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc 5941 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat 6001 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag 6061 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg 6121 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc 6181 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca 6241 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg 6301 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc 6361 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta 6421 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc 6481 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg 6541 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc 6601 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc 6661 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat 6721 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag 6781 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc 6841 ccgaaaagtg ccacctgacg tc //