LOCUS Exported 7146 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pGEN-mSmoA1 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7146) AUTHORS Taipale J, Chen JK, Cooper MK, Wang B, Mann RK, Milenkovic L, Scott MP, Beachy PA TITLE Effects of oncogenic mutations in Smoothened and Patched can be reversed by cyclopamine. JOURNAL Nature. 2000 Aug 31;406(6799):1005-9. PUBMED 10984056 REFERENCE 2 (bases 1 to 7146) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7146 /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" regulatory 2948..2957 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 2954..3670 /codon_start=1 /product="enhanced CFP" /label=ECFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(2999..3020) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(3260..3279) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 3607..3628 /label=EGFP-C /note="EGFP, forward primer" polyA_signal 3874..3995 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(3911..3930) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 3965..3984 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin complement(4002..4457) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(4139..4160) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 4351..4370 /label=F1ori-R /note="F1 origin, reverse primer" promoter 4484..4588 /gene="bla" /label=AmpR promoter promoter 4590..4947 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(4613..4633) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 4798..4933 /label=SV40 ori /note="SV40 origin of replication" primer_bind 4860..4879 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 4982..5776 /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(5036..5055) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 5646..5665 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(5964..5983) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 6008..6055 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 6384..6972 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 6873..6892 /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 gctacaagct 541 taccatggcc gctggccgcc ccgtgcgtgg gcccgagctg gcgccccgga ggctgctgca 601 gttgctgctg ctggtactgc tggggggccc gggccggggg gcggccttga gcgggaacgt 661 gaccgggcct gggcctcaca gcgccagcgg gagctcgagg aggaacgtgc cggtgaccag 721 ccctccgccg ccgctgctga gccactgcgg ccgggccgcc cactgcgagc ctttgcgcta 781 caacgtgtgc ctgggctcgg cgctgcccta cggagccacc accacgctgc tggctgggga 841 ctcggactcg caggaggaag cgcacggcaa gctcgtgctc tggtccggcc tccggaatgc 901 cccccgctgc tgggcagtga tccagcccct gctgtgtgct gtctacatgc ccaagtgtga 961 gaatgaccga gtggagttgc ccagccgtac cctctgccag gccacccgag gcccctgtgc 1021 cattgtggag cgggagcgag ggtggcctga ctttctgcgt tgcacaccgg accacttccc 1081 tgaaggctgc ccaaacgagg tacaaaacat caagttcaac agctcaggcc aatgtgaagc 1141 acccttggtg cgaacagaca accccaagag ctggtatgag gacgtggagg gctgtgggat 1201 tcagtgtcag aacccgctgt tcaccgaggc cgagcaccag gacatgcaca gctacatcgc 1261 agccttcggg gcggtcactg gtctctgcac gctcttcaca ctggccacct ttgtggctga 1321 ctggcggaat tccaatcgct accctgcggt tattctcttc tatgtcaatg cgtgtttctt 1381 cgtgggcagc attggctggc tggcccagtt catggatggt gcccgccgag agattgtttg 1441 ccgagcagat ggcaccatga gatttgggga gcccacctcc agtgagaccc tgtcctgtgt 1501 catcatcttt gtcattgtgt actatgcctt gatggctgga gtagtctggt tcgtggtcct 1561 cacctatgcc tggcacacct ccttcaaagc cctgggcacc acctaccagc ctctctcggg 1621 caagacatcc tatttccacc tgctcacgtg gtcactcccc tttgtcctca cggtggcaat 1681 cctggctgtg gctcaggtag atggagactc cgtgagtggc atctgttttg taggctacaa 1741 gaactatcgg taccgtgctg gctttgtcct ggccccaatt ggcctggtgc ttattgtggg 1801 aggctacttc ctcatcagag gggtcatgac tctgttctcc atcaagagca accaccctgg 1861 gctcttaagt gagaaggcag ccagcaagat caacgagacc atgctgcgcc tgggcatttt 1921 tggcttcctg gcctttggct ttgtgctcat caccttcagc tgccacttct atgacttctt 1981 caaccaggct gagtgggagc gtagcttccg ggactatgtg ctatgccaag ccaacgtgac 2041 catcgggctg cctaccaaga agcccattcc tgactgtgag atcaagaatc ggcccagcct 2101 cctggtggag aagatcaatc tatttgccat gtttggcact ggcattgcca tgagcaccct 2161 cgtgtggacc aaggccaccc tgctcatctg gaggcgcacc tggtgcaggt tgactgggca 2221 cagtgatgat gagcccaaga gaatcaagaa gagcaagatg atcgccaagg ccttctctaa 2281 gcggcgtgag ctgctgcaga acccgggcca ggagctctcc ttcagcatgc acactgtctc 2341 ccatgatgga cctgttgcgg gtttggcttt tgacctcaat gaaccctcag ctgacgtctc 2401 ctctgcctgg gctcagcatg tcaccaagat ggtggctcgg agaggagcca tattgcccca 2461 ggatgtgtcc gttacccctg tggcaactcc agtgccacca gaagaacaag ccaacatgtg 2521 gctggttgag gcagagatct ccccagagtt agagaagcgt ttgggccgga agaaaaagcg 2581 gaggaagagg aagaaggagg tgtgcccctt ggggccagcc cctgagcttc accactctgc 2641 ccctgttcct gccaccagtg cagttcctcg gctgcctcag ctgcctcggc agaagtgcct 2701 ggtagctgca aacgcctggg gaacagggga gtcctgccga cagggagcct ggactctagt 2761 ctccaacccc ttctgcccag agcctagtcc ccatcaagat ccatttctcc ctggtgcctc 2821 agccccccgg gtctgggctc agggccgcct ccaggggctg ggatccattc attcccgcac 2881 taacctaatg gaggctgaga tcttggatgc agactcggac ttcagaaaag agctagcgct 2941 accggtcgcc accatggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct 3001 ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg 3061 cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt 3121 gccctggccc accctcgtga ccaccctgac ctggggcgtg cagtgcttca gccgctaccc 3181 cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga 3241 gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga 3301 gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa 3361 catcctgggg cacaagctgg agtacaacta catcagccac aacgtctata tcaccgccga 3421 caagcagaag aacggcatca aggccaactt caagatccgc cacaacatcg aggacggcag 3481 cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct 3541 gcccgacaac cactacctga gcacccagtc cgccctgagc aaagacccca acgagaagcg 3601 cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga 3661 gctgtacaag tccggactca gatctcgagc tcaagcttcg aattctgcag gaattaggtc 3721 gtagtcgacg gtaccgcggg cccgggatcc accggatcta gataactgat cataatcagc 3781 cataccacat ttgtagaggt tttacttgct ttaaaaaacc tcccacacct ccccctgaac 3841 ctgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc ttataatggt 3901 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct 3961 agttgtggtt tgtccaaact catcaatgta tcttaacgcg taaattgtaa gcgttaatat 4021 tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga 4081 aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc 4141 agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac 4201 cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc 4261 gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg 4321 gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag 4381 ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc 4441 gccgctacag ggcgcgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg 4501 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat 4561 gcttcaataa tattgaaaaa ggaagagtcc tgaggcggaa agaaccagct gtggaatgtg 4621 tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg 4681 catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt 4741 atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc 4801 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt 4861 atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc 4921 ttttttggag gcctaggctt ttgcaaagat cgatcaagag acaggatgag gatcgtttcg 4981 catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt 5041 cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc 5101 agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact 5161 gcaagacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt 5221 gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca 5281 ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat 5341 gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg 5401 catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga 5461 agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcga gcatgcccga 5521 cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa 5581 tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga 5641 catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt 5701 cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct 5761 tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac 5821 ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc 5881 gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc 5941 gcccacccta gggggaggct aactgaaaca cggaaggaga caataccgga aggaacccgc 6001 gctatgacgg caataaaaag acagaataaa acgcacggtg ttgggtcgtt tgttcataaa 6061 cgcggggttc ggtcccaggg ctggcactct gtcgataccc caccgagacc ccattggggc 6121 caatacgccc gcgtttcttc cttttcccca ccccaccccc caagttcggg tgaaggccca 6181 gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag cctcaggtta ctcatatata 6241 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt 6301 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc 6361 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg 6421 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact 6481 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg 6541 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg 6601 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac 6661 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca 6721 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga 6781 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc 6841 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct 6901 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg 6961 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct 7021 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc 7081 atgcattagt tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga 7141 gttccg //