LOCUS V026595 7015 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V026595 VERSION V026595 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 874..1590 /codon_start=1 /note="mammalian codon-optimized" /product="the original enhanced GFP (Yang et al., 1996)" /transl_table=1 /translation="M,V,SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKL TLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFK DDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNG IKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMV LLEFVTAAGITLGMDELYK" /label="EGFP" CDS 1642..2505 /label="PDCD1(NM_005018)" /note="PDCD1(NM_005018)" /gene="PDCD1" polyA_signal 2582..2806 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 2852..3280 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 3294..3623 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3690..4484 /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 4658..4779 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4828..4844) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4852..4868 /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(4876..4906) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4921..4942 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5230..5815) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5986..6846) /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(6847..6951) /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 ggctagcgcc accatggtga gcaagggcga ggagctgttc 901 accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc 961 gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc 1021 accaccggca agctgcccgt gccctggccc accctcgtga ccaccctgac ctacggcgtg 1081 cagtgcttca gccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg 1141 cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc 1201 cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc 1261 gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac 1321 aacgtctata tcatggccga caagcagaag aacggcatca aggtgaactt caagatccgc 1381 cacaacatcg aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc 1441 ggcgacggcc ccgtgctgct gcccgacaac cactacctga gcacccagtc cgccctgagc 1501 aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg 1561 atcactctcg gcatggacga gctgtacaag ggaggcggag gtagtggagg agggggaagc 1621 ggaggcggag ggagtgaatt catgcagatc ccacaggcgc cctggccagt cgtctgggcg 1681 gtgctacaac tgggctggcg gccaggatgg ttcttagact ccccagacag gccctggaac 1741 ccccccacct tctccccagc cctgctcgtg gtgaccgaag gggacaacgc caccttcacc 1801 tgcagcttct ccaacacatc ggagagcttc gtgctaaact ggtaccgcat gagccccagc 1861 aaccagacgg acaagctggc cgccttcccc gaggaccgca gccagcccgg ccaggactgc 1921 cgcttccgtg tcacacaact gcccaacggg cgtgacttcc acatgagcgt ggtcagggcc 1981 cggcgcaatg acagcggcac ctacctctgt ggggccatct ccctggcccc caaggcgcag 2041 atcaaagaga gcctgcgggc agagctcagg gtgacagaga gaagggcaga agtgcccaca 2101 gcccacccca gcccctcacc caggccagcc ggccagttcc aaaccctggt ggttggtgtc 2161 gtgggcggcc tgctgggcag cctggtgctg ctagtctggg tcctggccgt catctgctcc 2221 cgggccgcac gagggacaat aggagccagg cgcaccggcc agcccctgaa ggaggacccc 2281 tcagccgtgc ctgtgttctc tgtggactat ggggagctgg atttccagtg gcgagagaag 2341 accccggagc cccccgtgcc ctgtgtccct gagcagacgg agtatgccac cattgtcttt 2401 cctagcggaa tgggcacctc atcccccgcc cgcaggggct cagctgacgg ccctcggagt 2461 gcccagccac tgaggcctga ggatggacac tgctcttggc ccctctgagg atcctgatat 2521 ccagcacagt ggcggccgct cgagtctaga gggcccgttt aaacccgctg atcagcctcg 2581 actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc 2641 ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt 2701 ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat 2761 tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa 2821 agaaccagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc attaagcgcg 2881 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct 2941 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta 3001 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa 3061 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct 3121 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc 3181 aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg 3241 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc 3301 agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc 3361 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc 3421 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc 3481 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt 3541 atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt 3601 ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt ttcggatctg 3661 atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt 3721 ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct 3781 gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga 3841 ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg 3901 ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact 3961 ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg 4021 agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct 4081 gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg 4141 gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt 4201 tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg 4261 cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc 4321 ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag 4381 agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt 4441 cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt 4501 cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc 4561 cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca 4621 gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa 4681 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca 4741 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac 4801 ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc 4861 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta 4921 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa 4981 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat 5041 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg 5101 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc 5161 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt 5221 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag 5281 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc 5341 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc 5401 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt 5461 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt 5521 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc 5581 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa 5641 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa 5701 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg 5761 tagcggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga 5821 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat 5881 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag 5941 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat 6001 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc 6061 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat 6121 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag 6181 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg 6241 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc 6301 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca 6361 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg 6421 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc 6481 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta 6541 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc 6601 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg 6661 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc 6721 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc 6781 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat 6841 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag 6901 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc 6961 ccgaaaagtg ccacctgacg tcgacggatc gggagatctc ccgatcccct atggt //