LOCUS Exported 5552 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pCMV-Tag4A-MITF-M (wt) SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5552) TITLE MITF in melanoma REFERENCE 2 (bases 1 to 5552) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5552 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 1..304 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 305..508 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 459..479 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" promoter 554..572 /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" primer_bind complement(1892..1908) /label=KS primer /note="common sequencing primer, one of multiple similar variants" primer_bind complement(1893..1909) /label=pBluescriptKS /note="For pBluescript vector" CDS 1911..1934 /codon_start=1 /product="FLAG(R) epitope tag, followed by an enterokinase cleavage site" /label=FLAG /translation="DYKDDDDK" primer_bind complement(1987..2006) /label=T7 /note="T7 promoter, forward primer" promoter complement(1988..2006) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" polyA_signal 2280..2401 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(2317..2336) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 2371..2390 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(2408..2863) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2545..2566) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 2757..2776 /label=F1ori-R /note="F1 origin, reverse primer" promoter 2890..2994 /gene="bla" /label=AmpR promoter promoter 2996..3353 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(3019..3039) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 3204..3339 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3266..3285 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 3388..4182 /codon_start=1 /gene="aph(3')-II (or nptII)" /product="aminoglycoside phosphotransferase from Tn5" /label=NeoR/KanR /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin(R))" /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(3442..3461) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 4052..4071 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(4370..4389) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 4414..4461 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 4790..5378 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 5279..5298 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt 61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca 121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc 181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta 241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac 301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg 361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg 421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt 481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgatt 541 acgccaagct cgaaattaac cctcactaaa gggaacaaaa gctggagctc caccgcggtg 601 gcggccgctc tagcccgggc ggatcccccg ggctgcagga attcatgctg gaaatgctag 661 aatataatca ctatcaggtg cagacccacc tcgaaaaccc caccaagtac cacatacagc 721 aagcccaacg gcagcaggta aagcagtacc tttctaccac tttagcaaat aaacatgcca 781 accaagtcct gagcttgcca tgtccaaacc agcctggcga tcatgtcatg ccaccggtgc 841 cggggagcag cgcacccaac agccccatgg ctatgcttac gcttaactcc aactgtgaaa 901 aagagggatt ttataagttt gaagagcaaa acagggcaga gagcgagtgc ccaggcatga 961 acacacattc acgagcgtcc tgtatgcaga tggatgatgt aatcgatgac atcattagcc 1021 tagaatcaag ttataatgag gaaatcttgg gcttgatgga tcctgctttg caaatggcaa 1081 atacgttgcc tgtctcggga aacttgattg atctttatgg aaaccaaggt ctgcccccac 1141 caggcctcac catcagcaac tcctgtccag ccaaccttcc caacataaaa agggagctca 1201 cagagtctga agcaagagca ctggccaaag agaggcagaa aaaggacaat cacaacctga 1261 ttgaacgaag aagaagattt aacataaatg accgcattaa agaactaggt actttgattc 1321 ccaagtcaaa tgatccagac atgcgctgga acaagggaac catcttaaaa gcatccgtgg 1381 actatatccg aaagttgcaa cgagaacagc aacgcgcaaa agaacttgaa aaccgacaga 1441 agaaactgga gcacgccaac cggcatttgt tgctcagaat acaggaactt gaaatgcagg 1501 ctcgagctca tggactttcc cttattccat ccacgggtct ctgctctcca gatttggtga 1561 atcggatcat caagcaagaa cccgttcttg agaactgcag ccaagacctc cttcagcatc 1621 atgcagacct aacctgtaca acaactctcg atctcacgga tggcaccatc accttcaaca 1681 acaacctcgg aactgggact gaggccaacc aagcctatag tgtccccaca aaaatgggat 1741 ccaaactgga agacatcctg atggacgaca ccctttctcc cgtcggtgtc actgatccac 1801 tcctttcctc agtgtccccc ggagcttcca aaacaagcag ccggaggagc agtatgagca 1861 tggaagagac ggagcacact tgtaagctta tcgataccgt cgacctcgag gattacaagg 1921 atgacgacga taagtagggc ccggtacctt aattaattaa ggtaccaggt aagtgtaccc 1981 aattcgccct atagtgagtc gtattacaat tcactcgatc gcccttccca acagttgcgc 2041 agcctgaatg gcgaatggag atccaatttt taagtgtata atgtgttaaa ctactgattc 2101 taattgtttg tgtattttag attcacagtc ccaaggctca tttcaggccc ctcagtcctc 2161 acagtctgtt catgatcata atcagccata ccacatttgt agaggtttta cttgctttaa 2221 aaaacctccc acacctcccc ctgaacctga aacataaaat gaatgcaatt gttgttgtta 2281 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa 2341 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt 2401 aacgcgtaaa ttgtaagcgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc 2461 agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag 2521 accgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg 2581 gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca 2641 tcaccctaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa 2701 gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg 2761 aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta 2821 accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtcaggtgg cacttttcgg 2881 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg 2941 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gaatcctgag 3001 gcggaaagaa ccagctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc 3061 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt 3121 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca 3181 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc 3241 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg 3301 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaagatcgat 3361 caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct 3421 ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc 3481 tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc 3541 gacctgtccg gtgccctgaa tgaactgcaa gacgaggcag cgcggctatc gtggctggcc 3601 acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg 3661 ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag 3721 aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc 3781 ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt 3841 cttgtcgatc aggatgatct ggacgaagaa catcaggggc tcgcgccagc cgaactgttc 3901 gccaggctca aggcgagcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc 3961 tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg 4021 ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagaa 4081 cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg 4141 cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg 4201 aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct 4261 tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc 4321 gcggggatct catgctggag ttcttcgccc accctagggg gaggctaact gaaacacgga 4381 aggagacaat accggaagga acccgcgcta tgacggcaat aaaaagacag aataaaacgc 4441 acggtgttgg gtcgtttgtt cataaacgcg gggttcggtc ccagggctgg cactctgtcg 4501 ataccccacc gagaccccat tggggccaat acgcccgcgt ttcttccttt tccccacccc 4561 accccccaag ttcgggtgaa ggcccagggc tcgcagccaa cgtcggggcg gcaggccctg 4621 ccatagcctc aggttactca tatatacttt agattgattt aaaacttcat ttttaattta 4681 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt 4741 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt 4801 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt 4861 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc 4921 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg 4981 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg 5041 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt 5101 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac 5161 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg 5221 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg 5281 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat 5341 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt 5401 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg 5461 attctgtgga taaccgtatt accgccatgc attagttatt aatagtaatc aattacgggg 5521 tcattagttc atagcccata tatggagttc cg //