LOCUS       Exported                4755 bp ds-DNA     circular SYN 09-SEP-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
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Genet."; date: "2009-06"; volume: "41(6)"; pages: "753-61. Epub 2009
            May 3"
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"