LOCUS V026876 7102 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V026876 VERSION V026876 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..2592 /label="LGALS8(NM_201543)" /note="LGALS8(NM_201543)" /gene="LGALS8" polyA_signal 2669..2893 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 2939..3367 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 3381..3710 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3777..4571 /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 4745..4866 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4915..4931) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4939..4955 /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(4963..4993) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 5008..5029 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5317..5902) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(6073..6933) /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(6934..7038) /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 catgatgttg tccttaaaca acctacagaa tatcatctat 1681 aacccggtaa tcccgtttgt tggcaccatt cctgatcagc tggatcctgg aactttgatt 1741 gtgatacgtg ggcatgttcc tagtgacgca gacagattcc aggtggatct gcagaatggc 1801 agcagcatga aacctcgagc cgatgtggcc tttcatttca atcctcgttt caaaagggcc 1861 ggctgcattg tttgcaatac tttgataaat gaaaaatggg gacgggaaga gatcacctat 1921 gacacgcctt tcaaaagaga aaagtctttt gagatcgtga ttatggtgct gaaggacaaa 1981 ttccaggtgg ctgtaaatgg aaaacatact ctgctctatg gccacaggat cggcccagag 2041 aaaatagaca ctctgggcat ttatggcaaa gtgaatattc actcaattgg ttttagcttc 2101 agctcggact tacaaagtac ccaagcatct agtctggaac tgacagagat aagtagagaa 2161 aatgttccaa agtctggcac gccccagctt aggctgccat tcgctgcaag gttgaacacc 2221 cccatgggcc ctggacgaac tgtcgtcgtt aaaggagaag tgaatgcaaa tgccaaaagc 2281 tttaatgttg acctactagc aggaaaatca aaggatattg ctctacactt gaacccacgc 2341 ctgaatatta aagcatttgt aagaaattct tttcttcagg agtcctgggg agaagaagag 2401 agaaatatta cctctttccc atttagtcct gggatgtact ttgagatgat aatttattgt 2461 gatgttagag aattcaaggt tgcagtaaat ggcgtacaca gcctggagta caaacacaga 2521 tttaaagagc tcagcagtat tgacacgctg gaaattaatg gagacatcca cttactggaa 2581 gtaaggagct ggtagggatc ctgatatcca gcacagtggc ggccgctcga gtctagaggg 2641 cccgtttaaa cccgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt 2701 tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa 2761 taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg 2821 gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg 2881 gtgggctcta tggcttctga ggcggaaaga accagctggg gctctagggg gtatccccac 2941 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct 3001 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg 3061 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt 3121 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca 3181 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga 3241 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa 3301 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac 3361 gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag 3421 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc 3481 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag 3541 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc 3601 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc 3661 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg 3721 gagcttgtat atccattttc ggatctgatc aagagacagg atgaggatcg tttcgcatga 3781 ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct 3841 atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc 3901 aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg 3961 acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg 4021 acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc 4081 tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc 4141 ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg 4201 agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc 4261 atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg 4321 aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc 4381 gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag 4441 cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg 4501 tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg 4561 agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc 4621 atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt 4681 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca 4741 ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt 4801 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt 4861 atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata 4921 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag 4981 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg 5041 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca 5101 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc 5161 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg 5221 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa 5281 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga 5341 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag 5401 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct 5461 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg 5521 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc 5581 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt 5641 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta 5701 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac 5761 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc 5821 ttgatccggc aaacaaacca ccgctggtag cggttttttt gtttgcaagc agcagattac 5881 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca 5941 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac 6001 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac 6061 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt 6121 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt 6181 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt 6241 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc 6301 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa 6361 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg 6421 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt 6481 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc 6541 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt 6601 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg 6661 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac 6721 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc 6781 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt 6841 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg 6901 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag 6961 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa 7021 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtcg acggatcggg 7081 agatctcccg atcccctatg gt //