LOCUS V024618 6738 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V024618 VERSION V024618 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 1660..2202 /label="ARF1(NM_001024226)" /note="ARF1(NM_001024226)" /gene="ARF1" polyA_signal 2305..2529 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 2575..3003 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 3017..3346 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3413..4207 /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 4381..4502 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4551..4567) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4575..4591 /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(4599..4629) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4644..4665 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(4953..5538) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5709..6569) /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(6570..6674) /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 tccggactca gatctcgagg cggcggaggg 1621 agtggaggag ggggaagcgg aggcggaggg agtaagctta tggggaacat cttcgccaac 1681 ctcttcaagg gcctttttgg caaaaaagaa atgcgcatcc ttatggtggg cctggatgct 1741 gcggggaaga ccacgatcct gtacaagctt aagctgggtg agatcgtgac caccattccc 1801 accataggct tcaacgtgga aaccgtggag tacaagaaca tcagcttcac tgtgtgggat 1861 gtgggtggcc aggacaagat ccggcccctg tggcgccact acttccagaa cacacaaggc 1921 ctgatcttcg tggtggacag caatgacaga gagcgtgtga acgaggcccg tgaggagctc 1981 atgagaatgc tggccgagga cgagctccgg gatgctgtcc tcctggtgtt cgccaacaag 2041 caggacctcc ccaacgccat gaatgcagct gagatcacag acaagctggg gctgcactcc 2101 ctacgccaca ggaactggta cattcaggcc acctgcgcca ccagcggtga cgggctctat 2161 gaaggactgg actggctgtc caatcagctc cggaaccaga agtgaggatc cactagtcca 2221 gtgtggtgga attctgcaga tatccagcac agtggcggcc gctcgagtct agagggcccg 2281 tttaaacccg ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc 2341 cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa 2401 atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg 2461 ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg 2521 gctctatggc ttctgaggcg gaaagaacca gctggggctc tagggggtat ccccacgcgc 2581 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac 2641 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg 2701 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt 2761 tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc 2821 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct 2881 tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga 2941 ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga 3001 attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg 3061 cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg 3121 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc 3181 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca 3241 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt 3301 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc 3361 ttgtatatcc attttcggat ctgatcaaga gacaggatga ggatcgtttc gcatgattga 3421 acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga 3481 ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg 3541 gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga 3601 ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt 3661 tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct 3721 gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct 3781 gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg 3841 agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca 3901 ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga 3961 tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt 4021 ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt 4081 ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct 4141 ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt 4201 cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca 4261 cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg 4321 gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc 4381 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca 4441 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct 4501 tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg 4561 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata 4621 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca 4681 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc 4741 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg 4801 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta 4861 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc 4921 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag 4981 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac 5041 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc 5101 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt 5161 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc 5221 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga 5281 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta 5341 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta 5401 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga 5461 tccggcaaac aaaccaccgc tggtagcggt ttttttgttt gcaagcagca gattacgcgc 5521 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg 5581 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag 5641 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg 5701 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt 5761 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca 5821 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca 5881 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc 5941 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt 6001 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg 6061 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc 6121 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg 6181 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga 6241 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga 6301 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta 6361 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg 6421 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact 6481 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata 6541 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt 6601 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa 6661 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg atcgggagat 6721 ctcccgatcc cctatggt //