LOCUS       40924_11601        4755 bp DNA     circular SYN 09-SEP-2021
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
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)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 4755)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat
            Genet."; date: "2009-06"; volume: "41(6)"; pages: "753-61. Epub 2009
            May 3"
COMMENT     SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..4755
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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     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"
     promoter        828..846
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             857..1876
                     /codon_start=1
                     /label=SB100X
                     /note="hyperactive variant of the Sleeping Beauty
                     transposase (Mates et al., 2009)"
                     /translation="MGKSKEISQDLRKRIVDLHKSGSSLGAISKRLAVPRSSVQTIVRK
                     YKHHGTTQPSYRSGRRRVLSPRDERTLVRKVQINPRTTAKDLVKMLEETGTKVSISTVK
                     RVLYRHNLKGHSARKKPLLQNRHKKARLRFATAHGDKDRTFWRNVLWSDETKIELFGHN
                     DHRYVWRKKGEACKPKNTIPTVKHGGGSIMLWGCFAAGGTGALHKIDGIMDAVQYVDIL
                     KQHLKTSVRKLKLGRKWVFQHDNDPKHTSKVVAKWLKDNKVKVLEWPSQSPDLNPIENL
                     WAELKKRVRARRPTNLTQLHQLCQEEWAKIHPNYCGKLVEGYPKRLTQVKQFKGNATKY
                     "
     polyA_signal    complement(1896..2030)
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2141..2157)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(2165..2181)
                     /label=lac operator
                     /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    complement(2234..2255)
                     /label=CAP binding site
                     /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"
     CDS             complement(3863..4519)
                     /codon_start=1
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     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
//
