LOCUS V031741 6652 bp DNA circular SYN 01-JAN-1980 DEFINITION synthetic circular DNA. ACCESSION V031741 VERSION V031741 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 917..1441 /label="ARF6(NM_001663)" /note="ARF6(NM_001663)" /gene="ARF6" CDS 1466..2176 /codon_start=1 /note="mammalian codon-optimized" /product="monomeric derivative of DsRed fluorescent protein (Shaner et al., 2004)" /transl_table=1 /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA EGRHSTGGMDELYK*" /label="mCherry" promoter complement(2204..2222) /note="promoter for bacteriophage SP6 RNA polymerase" /label="SP6 promoter" polyA_signal 2248..2472 /note="bovine growth hormone polyadenylation signal" /label="bGH poly(A) signal" rep_origin 2518..2946 /direction=RIGHT /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /label="f1 ori" promoter 2960..3290 /note="SV40 enhancer and early promoter" /label="SV40 promoter" CDS 3357..4151 /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 4325..4446 /note="SV40 polyadenylation signal" /label="SV40 poly(A) signal" primer_bind complement(4495..4511) /note="common sequencing primer, one of multiple similar variants" /label="M13 rev" protein_bind 4519..4535 /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(4543..4573) /note="promoter for the E. coli lac operon" /label="lac promoter" protein_bind 4588..4609 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." /label="CAP binding site" rep_origin complement(4897..5485) /direction=LEFT /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" /label="ori" CDS complement(5656..6516) /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(6517..6621) /gene="bla" /label="AmpR promoter" ORIGIN 1 gacggatcgg gagatctccc gatcccctat ggtcgactct 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 atgggtggac 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 gcttggtacc 901 gagctcggat gccaccatgg ggaaggtgct atccaaaatc ttcgggaaca aggaaatgcg 961 gatcctcatg ttgggcctgg acgcggccgg caagacaaca atcctgtaca agttgaagct 1021 gggccagtcg gtgaccacca ttcccactgt gggtttcaac gtggagacgg tgacttacaa 1081 aaatgtcaag ttcaacgtat gggatgtggg cggccaggac aagatccggc cgctctggcg 1141 gcattactac actgggaccc aaggtctcat cttcgtagtg gactgcgccg accgcgaccg 1201 catcgatgag gctcgccagg agctgcaccg cattatcaat gaccgggaga tgagggacgc 1261 cataatcctc atcttcgcca acaagcagga cctgcccgat gccatgaaac cccacgagat 1321 ccaggagaaa ctgggcctga cccggattcg ggacaggaac tggtatgtgc agccctcctg 1381 tgccacctca ggggacggac tctatgaggg gctcacatgg ttaacctcta actacaaatc 1441 tctcgaggga cttaggtctg ccaccatggt gagcaagggc gaggaggata acatggccat 1501 catcaaggag ttcatgcgct tcaaggtgca catggagggc tccgtgaacg gccacgagtt 1561 cgagatcgag ggcgagggcg agggccgccc ctacgagggc acccagaccg ccaagctgaa 1621 ggtgaccaag ggtggccccc tgcccttcgc ctgggacatc ctgtcccctc agttcatgta 1681 cggctccaag gcctacgtga agcaccccgc cgacatcccc gactacttga agctgtcctt 1741 ccccgagggc ttcaagtggg agcgcgtgat gaacttcgag gacggcggcg tggtgaccgt 1801 gacccaggac tcctccctgc aggacggcga gttcatctac aaggtgaagc tgcgcggcac 1861 caacttcccc tccgacggcc ccgtaatgca gaagaagacc atgggctggg aggcctcctc 1921 cgagcggatg taccccgagg acggcgccct gaagggcgag atcaagcaga ggctgaagct 1981 gaaggacggc ggccactacg acgctgaggt caagaccacc tacaaggcca agaagcccgt 2041 gcagctgccc ggcgcctaca acgtcaacat caagttggac atcacctccc acaacgagga 2101 ctacaccatc gtggaacagt acgaacgcgc cgagggccgc cactccaccg gcggcatgga 2161 cgagctgtac aagtaactcg agcatgcatc tagagggccc tattctatag tgtcacctaa 2221 atgctagagc tcgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt 2281 gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat 2341 aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg 2401 tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg 2461 tgggctctat ggcttctgag gcggaaagaa ccagctgggg ctctaggggg tatccccacg 2521 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta 2581 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt 2641 tcgccggctt tccccgtcaa gctctaaatc ggggcatccc tttagggttc cgatttagtg 2701 ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat 2761 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac 2821 tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag 2881 ggattttggg gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg 2941 cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagg 3001 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc 3061 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag 3121 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc 3181 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc 3241 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg 3301 gagcttgtat atccattttc ggatctgatc aagagacagg atgaggatcg tttcgcatga 3361 ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct 3421 atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc 3481 aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg 3541 acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg 3601 acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc 3661 tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc 3721 ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg 3781 agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc 3841 atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg 3901 aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc 3961 gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag 4021 cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg 4081 tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg 4141 agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc 4201 atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt 4261 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca 4321 ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt 4381 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt 4441 atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata 4501 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag 4561 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg 4621 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca 4681 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc 4741 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg 4801 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa 4861 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga 4921 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag 4981 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct 5041 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc aatgctcacg 5101 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc 5161 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt 5221 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta 5281 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac 5341 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc 5401 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat 5461 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc 5521 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt 5581 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta 5641 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct 5701 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg 5761 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga 5821 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt 5881 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt 5941 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt 6001 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat 6061 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc 6121 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc 6181 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat 6241 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag 6301 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt 6361 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc 6421 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa 6481 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg 6541 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa 6601 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tc //