LOCUS Exported 4755 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pCMV(CAT)T7-SB100 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4755) AUTHORS Mates L, Chuah MK, Belay E, Jerchow B, Manoj N, Acosta-Sanchez A, Grzela DP, Schmitt A, Becker K, Matrai J, Ma L, Samara-Kuko E, Gysemans C, Pryputniewicz D, Miskey C, Fletcher B, VandenDriessche T, Ivics Z, Izsvak Z TITLE Molecular evolution of a novel hyperactive Sleeping Beauty transposase enables robust stable gene transfer in vertebrates. JOURNAL Nat Genet. 2009 Jun;41(6):753-61. Epub 2009 May 3. PUBMED 19412179 REFERENCE 2 (bases 1 to 4755) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4755 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 30..333 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 334..537 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 488..508 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 534..558 /label=LNCX /note="Human CMV promoter, forward primer" intron 675..771 /label=SV40 intron /note="modified SV40 intron with splice donor and acceptor sites" primer_bind 828..847 /label=T7 /note="T7 promoter, forward primer" promoter 828..846 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" CDS 857..1879 /codon_start=1 /product="hyperactive variant of the Sleeping Beauty transposase (Mates et al., 2009)" /label=SB100X /translation="MGKSKEISQDLRKRIVDLHKSGSSLGAISKRLAVPRSSVQTIVRK YKHHGTTQPSYRSGRRRVLSPRDERTLVRKVQINPRTTAKDLVKMLEETGTKVSISTVK RVLYRHNLKGHSARKKPLLQNRHKKARLRFATAHGDKDRTFWRNVLWSDETKIELFGHN DHRYVWRKKGEACKPKNTIPTVKHGGGSIMLWGCFAAGGTGALHKIDGIMDAVQYVDIL KQHLKTSVRKLKLGRKWVFQHDNDPKHTSKVVAKWLKDNKVKVLEWPSQSPDLNPIENL WAELKKRVRARRPTNLTQLHQLCQEEWAKIHPNYCGKLVEGYPKRLTQVKQFKGNATKY " polyA_signal 1896..2030 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(1920..1939) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 1974..1993 /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind complement(2141..2157) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(2141..2157) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(2154..2176) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 2165..2181 /label=lac operator /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-beta-D-thiogalactopyranoside (IPTG)." promoter complement(2189..2219) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 2234..2255 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(2372..2389) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(2543..3131) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(2623..2642) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(3860..4519) /codon_start=1 /gene="cat" /product="chloramphenicol acetyltransferase" /label=CmR /note="confers resistance to chloramphenicol" /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA" primer_bind 4434..4453 /label=CAT-R /note="Chloramphenicol resistance gene, reverse primer" promoter complement(4520..4622) /label=cat promoter /note="promoter of the E. coli cat gene encoding chloramphenicol acetyltransferase" ORIGIN 1 ggccaattcg agcttgcatg cctgcaggtc gttacataac ttacggtaaa tggcccgcct 61 ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta 121 acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac 181 ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt 241 aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag 301 tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat 361 gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat 421 gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc 481 ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctcgt 541 ttagtgaacc gtcagatcgc ctggagacgc catccacgct gttttgacct ccatagaaga 601 caccgggacc gatccagcct ccggactcta gaggatccgg tactcgagga actgaaaaac 661 cagaaagtta actggtaagt ttagtctttt tgtcttttat ttcaggtccc ggatccggtg 721 gtggtgcaaa tcaaagaact gctcctcagt ggatgttgcc tttacttcta ggcctgtacg 781 gaagtgttac ttctgctcta aaagctgcgg aattgtaccc gcggaattaa tacgactcac 841 tatagggact agtaccatgg gaaaatcaaa agaaatcagc caagacctca gaaaaagaat 901 tgtagacctc cacaagtctg gttcatcctt gggagcaatt tccaaacgcc tggcggtacc 961 acgttcatct gtacaaacaa tagtacgcaa gtataaacac catgggacca cgcagccgtc 1021 ataccgctca ggaaggagac gcgttctgtc tcctagagat gaacgtactt tggtgcgaaa 1081 agtgcaaatc aatcccagaa caacagcaaa ggaccttgtg aagatgctgg aggaaacagg 1141 tacaaaagta tctatatcca cagtaaaacg agtcctatat cgacataacc tgaaaggcca 1201 ctcagcaagg aagaagccac tgctccaaaa ccgacataag aaagccagac tacggtttgc 1261 aactgcacat ggggacaaag atcgtacttt ttggagaaat gtcctctggt ctgatgaaac 1321 aaaaatagaa ctgtttggcc ataatgacca tcgttatgtt tggaggaaga agggggaggc 1381 ttgcaagccg aagaacacca tcccaaccgt gaagcacggg ggtggcagca tcatgttgtg 1441 ggggtgcttt gctgcaggag ggactggtgc acttcacaaa atagatggca tcatggacgc 1501 ggtgcagtat gtggatatat tgaagcaaca tctcaagaca tcagtcagga agttaaagct 1561 tggtcgcaaa tgggtcttcc aacacgacaa tgaccccaag catacttcca aagttgtggc 1621 aaaatggctt aaggacaaca aagtcaaggt attggagtgg ccatcacaaa gccctgacct 1681 caatcctata gaaaatttgt gggcagaact gaaaaagcgt gtgcgagcaa ggaggcctac 1741 aaacctgact cagttacacc agctctgtca ggaggaatgg gccaaaattc acccaaatta 1801 ttgtgggaag cttgtggaag gctacccgaa acgtttgacc caagttaaac aatttaaagg 1861 caatgctacc aaatactagg ggccctaacc gcggggatcc agacatgata agatacattg 1921 atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt 1981 gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca 2041 attgcattca ttttatgttt caggttcagg gggaggtgtg ggaggttttt tcggatcctc 2101 tagagtcgac ctgcaggcat gcaagcttgg cgtaatcatg gtcatagctg tttcctgtgt 2161 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag 2221 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt 2281 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag 2341 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg 2401 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat 2461 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta 2521 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa 2581 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc 2641 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt 2701 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca 2761 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg 2821 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat 2881 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta 2941 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct 3001 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac 3061 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa 3121 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa 3181 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt 3241 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca 3301 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca 3361 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc 3421 ccagtgctgc aatgataccg tggatcgatc caccataccc acgccgaaac aagcgccctg 3481 caccattatg ttccggatct gcatcgcagg atgctgctgg ctaccctgtg gaacacctac 3541 atctgtatta acgaagcgct aaccgttttt atcaggctct gggaggcaga ataaatgatc 3601 atatcgtcaa ttattacctc cacggggaga gcctgagcaa actggcctca ggcatttgag 3661 aagcacacgg tcacactgct tccggtagtc aataaaccgg taaaccagca atagacataa 3721 gcggctattt aacgaccctg ccctgaaccg acgaccgggt cgaatttgct ttcgaatttc 3781 tgccattcat ccgcttatta tcacttattc aggcgtagca accaggcgtt taagggcacc 3841 aataactgcc ttaaaaaaat tacgccccgc cctgccactc atcgcagtac tgttgtaatt 3901 cattaagcat tctgccgaca tggaagccat cacaaacggc atgatgaacc tgaatcgcca 3961 gcggcatcag caccttgtcg ccttgcgtat aatatttgcc catggtgaaa acgggggcga 4021 agaagttgtc catattggcc acgtttaaat caaaactggt gaaactcacc cagggattgg 4081 ctgagacgaa aaacatattc tcaataaacc ctttagggaa ataggccagg ttttcaccgt 4141 aacacgccac atcttgcgaa tatatgtgta gaaactgccg gaaatcgtcg tggtattcac 4201 tccagagcga tgaaaacgtt tcagtttgct catggaaaac ggtgtaacaa gggtgaacac 4261 tatcccatat caccagctca ccgtctttca ttgccatacg gaattccgga tgagcattca 4321 tcaggcgggc aagaatgtga ataaaggccg gataaaactt gtgcttattt ttctttacgg 4381 tctttaaaaa ggccgtaata tccagctgaa cggtctggtt ataggtacat tgagcaactg 4441 actgaaatgc ctcaaaatgt tctttacgat gccattggga tatatcaacg gtggtatatc 4501 cagtgatttt tttctccatt ttagcttcct tagctcctga aaatctcgat aactcaaaaa 4561 atacgcccgg tagtgatctt atttcattat ggtgaaagtt ggaacctctt acgtgccgat 4621 caacgtctca ttttcgccaa aagttggccc agggcttccc ggtatcaaca gggacaccag 4681 gatttattta ttctgcgaag tgatcttccg tcacaggtat ttattcggcg caaagtgcgt 4741 cgggtcacta cgtga //