LOCUS V047508 6468 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V047508 VERSION V047508 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..1941 /label="HMGN1(NM_004965)" /note="HMGN1(NM_004965)" /gene="HMGN1" CDS 1942..1959 /codon_start=1 /product="6xHis affinity tag" /transl_table=1 /translation="HHHHHH" /label="6xHis" polyA_signal 2035..2259 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 2305..2733 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 2747..3076 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3143..3937 /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 4111..4232 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4281..4297) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4305..4321 /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(4329..4359) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4374..4395 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(4683..5268) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5439..6299) /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(6300..6404) /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 catgcccaag aggaaggtca gctccgccga aggcgccgcc 1681 aaggaagagc ccaagaggag atcggcgcgg ttgtcagcta aacctcctgc aaaagtggaa 1741 gcgaagccga aaaaggcagc agcgaaggat aaatcttcag acaaaaaagt gcaaacaaaa 1801 gggaaaaggg gagcaaaggg aaaacaggcc gaagtggcta accaagaaac taaagaagac 1861 ttacctgcgg aaaacgggga aacgaagact gaggagagtc cagcctctga tgaagcagga 1921 gagaaagaag ccaagtctga tcatcatcac catcaccatt aggatcctga tatccagcac 1981 agtggcggcc gctcgagtct agagggcccg tttaaacccg ctgatcagcc tcgactgtgc 2041 cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag 2101 gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta 2161 ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag 2221 acaatagcag gcatgctggg gatgcggtgg gctctatggc ttctgaggcg gaaagaacca 2281 gctggggctc tagggggtat ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg 2341 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg 2401 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg 2461 ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt 2521 agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt 2581 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta 2641 tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa 2701 atgagctgat ttaacaaaaa tttaacgcga attaattctg tggaatgtgt gtcagttagg 2761 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta 2821 gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat 2881 gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac 2941 tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga 3001 ggccgaggcc gcctctgcct ctgagctatt ccagaagtag tgaggaggct tttttggagg 3061 cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc attttcggat ctgatcaaga 3121 gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc 3181 cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga 3241 tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct 3301 gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac 3361 gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct 3421 attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt 3481 atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt 3541 cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt 3601 cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag 3661 gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt 3721 gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg 3781 tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg 3841 cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg 3901 catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg 3961 accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat 4021 gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg 4081 gatctcatgc tggagttctt cgcccacccc aacttgttta ttgcagctta taatggttac 4141 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt 4201 tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtataccgtc gacctctagc 4261 tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca 4321 attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg 4381 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg 4441 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc 4501 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta 4561 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag 4621 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg 4681 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg 4741 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg 4801 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga 4861 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc 4921 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt 4981 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact 5041 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg 5101 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt 5161 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt 5221 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg 5281 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc 5341 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa 5401 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag 5461 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg 5521 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga 5581 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag 5641 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa 5701 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc 5761 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca 5821 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg 5881 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat 5941 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc 6001 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg 6061 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg 6121 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt 6181 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca 6241 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata 6301 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac 6361 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa 6421 gtgccacctg acgtcgacgg atcgggagat ctcccgatcc cctatggt //