LOCUS V032536 8902 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V032536 VERSION V032536 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..4392 /label="HK1(NM_000188)" /note="HK1(NM_000188)" /gene="HK1" polyA_signal 4469..4693 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 4739..5167 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 5181..5510 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 5577..6371 /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 6545..6666 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(6715..6731) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 6739..6755 /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(6763..6793) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 6808..6829 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(7117..7702) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(7873..8733) /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(8734..8838) /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 catgatcgcc gcgcagctcc tggcctatta cttcacggag 1681 ctgaaggatg accaggtcaa aaagattgac aagtatctct atgccatgcg gctctccgat 1741 gaaactctca tagatatcat gactcgcttc aggaaggaga tgaagaatgg cctctcccgg 1801 gattttaatc caacagccac agtcaagatg ttgccaacat tcgtaaggtc cattcctgat 1861 ggctctgaaa agggagattt cattgccctg gatcttggtg ggtcttcctt tcgaattctg 1921 cgggtgcaag tgaatcatga gaaaaaccag aatgttcaca tggagtccga ggtttatgac 1981 accccagaga acatcgtgca cggcagtgga agccagcttt ttgatcatgt tgctgagtgc 2041 ctgggagatt tcatggagaa aaggaagatc aaggacaaga agttacctgt gggattcacg 2101 ttttcttttc cttgccaaca atccaaaata gatgaggcca tcctgatcac ctggacaaag 2161 cgatttaaag cgagcggagt ggaaggagca gatgtggtca aactgcttaa caaagccatc 2221 aaaaagcgag gggactatga tgccaacatc gtagctgtgg tgaatgacac agtgggcacc 2281 atgatgacct gtggctatga cgaccagcac tgtgaagtcg gcctgatcat cggcactggc 2341 accaatgctt gctacatgga ggaactgagg cacattgatc tggtggaagg agacgagggg 2401 aggatgtgta tcaatacaga atggggagcc tttggagacg atggatcatt agaagacatc 2461 cggacagagt ttgacaggga gatagaccgg ggatccctca accctggaaa acagctgttt 2521 gagaagatgg tcagtggcat gtacttggga gagctggttc gactgatcct agtcaagatg 2581 gccaaggagg gcctcttatt tgaagggcgg atcaccccgg agctgctcac ccgagggaag 2641 tttaacacca gtgatgtgtc agccatcgaa aagaataagg aaggcctcca caatgccaaa 2701 gaaatcctga cccgcctggg agtggagccg tccgatgatg actgtgtctc agtccagcac 2761 gtttgcacca ttgtctcatt tcgctcagcc aacttggtgg ctgccacact gggcgccatc 2821 ttgaaccgcc tgcgtgataa caagggcaca cccaggctgc ggaccacggt tggtgtcgac 2881 ggatctcttt acaagacgca cccacagtat tcccggcgtt tccacaagac tctaaggcgc 2941 ttggtgccag actccgatgt gcgcttcctc ctctcggaga gtggcagcgg caagggggct 3001 gccatggtga cggcggtggc ctaccgcttg gccgagcagc accggcagat agaggagacc 3061 ctggctcatt tccacctcac caaggacatg ctgctggagg tgaagaagag gatgcgggcc 3121 gagatggagc tggggctgag gaagcagacg cacaacaatg ccgtggttaa gatgctgccc 3181 tccttcgtcc ggagaactcc cgacgggacc gagaatggtg acttcttggc cctggatctt 3241 ggaggaacca atttccgtgt gctgctggtg aaaatccgta gtgggaaaaa gagaacggtg 3301 gaaatgcaca acaagatcta cgccattcct attgaaatca tgcagggcac tggggaagag 3361 ctgtttgatc acattgtctc ctgcatctct gacttcttgg actacatggg gatcaaaggc 3421 cccaggatgc ctctgggctt cacgttctca tttccctgcc agcagacgag tctggacgcg 3481 ggaatcttga tcacgtggac aaagggtttt aaggcaacag actgcgtggg ccacgatgta 3541 gtcaccttac taagggatgc gataaaaagg agagaggaat ttgacctgga cgtggtggct 3601 gtggtcaacg acacagtggg caccatgatg acctgtgctt atgaggagcc cacctgtgag 3661 gttggactca ttgttgggac cggcagcaat gcctgctaca tggaggagat gaagaacgtg 3721 gagatggtgg agggggacca ggggcagatg tgcatcaaca tggagtgggg ggcctttggg 3781 gacaacgggt gtctggatga tatcaggaca cactacgaca gactggtgga cgaatattcc 3841 ctaaatgctg ggaaacaaag gtatgagaag atgatcagtg gtatgtacct gggtgaaatc 3901 gtccgcaaca tcttaatcga cttcaccaag aagggattcc tcttccgagg gcagatctct 3961 gagacgctga agacccgggg catctttgag accaagtttc tctctcagat cgagagtgac 4021 cgattagcac tgctccaggt ccgggctatc ctccagcagc taggtctgaa tagcacctgc 4081 gatgacagta tcctcgtcaa gacagtgtgc ggggtggtgt ccaggagggc cgcacagctg 4141 tgtggcgcag gcatggctgc ggttgtggat aagatccgcg agaacagagg actggaccgt 4201 ctgaatgtga ctgtgggagt ggacgggaca ctctacaagc ttcatccaca cttctccaga 4261 atcatgcacc agacggtgaa ggaactgtca ccaaaatgta acgtgtcctt cctcctgtct 4321 gaggatggca gcggcaaggg ggccgccctc atcacggccg tgggcgtgcg gttacgcaca 4381 gaggcaagca gctaaggatc ctgatatcca gcacagtggc ggccgctcga gtctagaggg 4441 cccgtttaaa cccgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt 4501 tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa 4561 taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg 4621 gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg 4681 gtgggctcta tggcttctga ggcggaaaga accagctggg gctctagggg gtatccccac 4741 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct 4801 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg 4861 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt 4921 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca 4981 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga 5041 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa 5101 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac 5161 gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag 5221 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc 5281 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag 5341 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc 5401 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc 5461 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg 5521 gagcttgtat atccattttc ggatctgatc aagagacagg atgaggatcg tttcgcatga 5581 ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct 5641 atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc 5701 aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg 5761 acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg 5821 acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc 5881 tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc 5941 ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg 6001 agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc 6061 atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg 6121 aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc 6181 gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag 6241 cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg 6301 tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg 6361 agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc 6421 atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt 6481 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca 6541 ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt 6601 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt 6661 atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata 6721 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag 6781 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg 6841 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca 6901 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc 6961 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg 7021 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa 7081 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga 7141 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag 7201 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct 7261 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg 7321 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc 7381 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt 7441 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta 7501 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac 7561 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc 7621 ttgatccggc aaacaaacca ccgctggtag cggttttttt gtttgcaagc agcagattac 7681 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca 7741 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac 7801 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac 7861 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt 7921 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt 7981 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt 8041 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc 8101 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa 8161 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg 8221 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt 8281 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc 8341 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt 8401 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg 8461 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac 8521 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc 8581 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt 8641 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg 8701 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag 8761 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa 8821 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtcg acggatcggg 8881 agatctcccg atcccctatg gt //