LOCUS V038959 7055 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V038959 VERSION V038959 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 923..1639 /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 1676..2545 /label="Dlk1(NM_001190703)" /note="Dlk1(NM_001190703)" /gene="Dlk1" CDS 2552..2617 /codon_start=1 /product="three tandem FLAG® epitope tags, followed by an enterokinase cleavage site" /transl_table=1 /translation="DYKDHDGDYKDHDIDYKDDDDK" /label="3xFLAG" polyA_signal 2655..2879 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 2925..3353 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 3367..3696 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3763..4557 /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 4731..4852 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4901..4917) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4925..4941 /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(4949..4979) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4994..5015 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(5303..5888) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(6059..6919) /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(6920..7024) /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 aagcttgcca ccatggtgag caagggcgag gagctgttca ccggggtggt 961 gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga 1021 gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa 1081 gctgcccgtg ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag 1141 ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta 1201 cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt 1261 gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga 1321 ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat 1381 catggccgac aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga 1441 ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc 1501 cgtgctgctg cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa 1561 cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg 1621 catggacgag ctgtacaagg gcggtggagg cagtggtgga ggcggtagcg aattcatgat 1681 cgcgaccgga gccctcctgc gcgtcctctt gctcctgctg gctttcggcc acagcaccta 1741 tggggctgaa tgcgacccac cctgtgaccc ccagtatgga ttctgcgagg ctgacaatgt 1801 ctgcaggtgc catgttggct gggagggtcc cctctgtgac aagtgtgtaa ctgcccctgg 1861 ctgtgtcaat ggagtctgca aggaaccatg gcagtgcatc tgcaaggatg gctgggacgg 1921 gaaattctgc gaaatagacg ttcgggcttg cacctcaacc ccctgcgcca acaatggaac 1981 ttgcgtggac ctggagaaag gccagtacga atgctcctgc acacctgggt tctctggaaa 2041 ggactgccag cacaaggctg ggccctgcgt gatcaatggt tctccctgcc agcacggagg 2101 cgcctgcgtg gatgatgagg gccaggcctc gcatgcttcc tgcctgtgcc cccctggctt 2161 ctcaggcaac ttctgtgaga tcgtagccgc aaccaacagc tgtaccccta acccatgcga 2221 gaacgatggc gtctgcaccg acatcggggg tgacttccgt tgccgctgcc cagctggatt 2281 cgtcgacaag acctgcagcc gcccggccat ctgcttcacc atcctgggcg tgctcaccag 2341 cctggtggtg ctgggcaccg tggccatcgt ctttctcaac aagtgcgaaa cctgggtgtc 2401 caacctgcgc tacaaccaca tgcttcgcaa gaagaagaac ctcctgttgc agtataacag 2461 cggcgaggag ctggcggtca atatcatctt ccccgagaag attgacatga ccactttcaa 2521 caaggaggct ggtgatgagg agatcctcga ggactacaaa gaccatgacg gtgattataa 2581 agatcatgac atcgactaca aggatgacga tgacaagtag tgagggcccg tttaaacccg 2641 ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt 2701 gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat 2761 tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag 2821 caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc 2881 ttctgaggcg gaaagaacca gctggggctc tagggggtat ccccacgcgc cctgtagcgg 2941 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc 3001 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc 3061 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct 3121 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac 3181 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac 3241 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat 3301 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg 3361 tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg 3421 caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca 3481 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact 3541 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta 3601 atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag 3661 tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc 3721 attttcggat ctgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga 3781 ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa 3841 cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt 3901 ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg 3961 ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa 4021 gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac 4081 cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt 4141 gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact 4201 cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg 4261 ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg 4321 acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc 4381 atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt 4441 gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc 4501 gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg 4561 ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg 4621 attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct 4681 ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta 4741 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat 4801 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct 4861 gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt 4921 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag 4981 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt 5041 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag 5101 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg 5161 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat 5221 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta 5281 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa 5341 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc 5401 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt 5461 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca 5521 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg 5581 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat 5641 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta 5701 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct 5761 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac 5821 aaaccaccgc tggtagcggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag 5881 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact 5941 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa 6001 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt 6061 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag 6121 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca 6181 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc 6241 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt 6301 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg 6361 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca 6421 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg 6481 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca 6541 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg 6601 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct 6661 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca 6721 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca 6781 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg 6841 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac 6901 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt 6961 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc 7021 cgcgcacatt tccccgaaaa gtgccacctg acgtc //