LOCUS       Exported                5725 bp DNA     circular SYN 15-OCT-2025
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5725)
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 5725)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5725
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             48..275
                     /label=IR/DR(R)
                     /note="Inverted/direct repeats of Sleeping Beauty 
                     transposon (Right)"
     promoter        complement(336..354)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(375..391)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(399..415)
                     /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(423..453)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(468..489)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(777..1365)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(1537..2397)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAAKLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(2398..2502)
                     /label=AmpR promoter
     rep_origin      2529..2984
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     primer_bind     3125..3141
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        3151..3169
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     LTR             3195..3421
                     /label=IR/DR(L)
                     /note="Inverted/direct repeats of Sleeping Beauty 
                     transposon (Left)"
     enhancer        3588..3891
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3893..4170
                     /label=chicken beta-actin promoter
     CDS             4684..5349
                     /codon_start=1
                     /label=CopGFP
                     /note="green fluorescent protein 2 from Pontellina plumata,
                     also known as ppluGFP2 (Shagin et al., 2004)"
                     /translation="MPAMKIECRITGTLNGVEFELVGGGEGTPEQGRMTNKMKSTKGAL
                     TFSPYLLSHVMGYGFYHFGTYPSGYENPFLHAINNGGYTNTRIEKYEDGGVLHVSFSYR
                     YEAGRVIGDFKVVGTGFPEDSVIFTDKIIRSNATVEHLHPMGDNVLVGSFARTFSLRDG
                     GYYSFVVDSHMHFKSAIHPSILQNGGPMFAFRRVEELHSNTELGIVEYQHAFKTPIAFA
                     "
     polyA_signal    complement(5392..5473)
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
ORIGIN
        1 ccgaatttgc cagttaacaa tttaaaggca atgctaccaa atactaattg agtgtatgta
       61 aacttctgac ccactgggaa tgtgatgaaa gaaataaaag ctgaaatgaa tcattctctc
      121 tactattatt ctgatatttc acattcttaa aataaagtgg tgatcctaac tgacctaaga
      181 cagggaattt ttactaggat taaatgtcag gaattgtgaa aaagtgagtt taaatgtatt
      241 tggctaaggt gtatgtaaac ttccgacttc aactgtatag ggatcctcta gctagagtcg
      301 acctcgaggg ggggcccggt acccagcttt tgttcccttt agtgagggtt aattgcgcgc
      361 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
      421 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
      481 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
      541 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
      601 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
      661 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
      721 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
      781 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
      841 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
      901 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaaccgtg
      961 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     1021 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     1081 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     1141 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     1201 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     1261 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     1321 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     1381 ttttttacgg ggtcttgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     1441 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     1501 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     1561 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     1621 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     1681 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     1741 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     1801 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     1861 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     1921 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     1981 cgttgtcaga agtaacttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     2041 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     2101 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     2161 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     2221 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     2281 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     2341 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     2401 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     2461 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     2521 gccacctgac gcgccctgta gcggcgcatt aagcgcggcg ggagtggtgg ttacgcgcag
     2581 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
     2641 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
     2701 ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg
     2761 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     2821 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt
     2881 tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca
     2941 aaaatttaac gcgaatttta acaaaatatt aacgcttaca atttccattc gccattcagg
     3001 ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg ccagctggcg
     3061 aaagggggat gtgctgcaag gcgattaagt tgggtaacgc cagggttttc ccagtcacga
     3121 cgttgtaaaa cgacggccag tgagcgcgcg taatacgact cactataggg cgaattggag
     3181 ctcggatccc tatacagttg aagtcggaag tttacataca cttaagttgg agtcattaaa
     3241 actcgttttt caactactcc acaaatttct tgttaacaaa caatagtttt ggcaagtcag
     3301 ttaggacatc tactttgtgc atgacacaag tcatttttcc aacaattgtt tacagacaga
     3361 ttatttcact tataattcac tgtatcacaa ttccagtggg tcagaagttt acatacacta
     3421 agttgactgt gcctttaaac agcttggaaa attccagaaa atgatgtcat ggctttagaa
     3481 gcttgatatc catggaattc actagtgcgc gcggccgctc tagcccctag ttattaatag
     3541 taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt
     3601 acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg
     3661 acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat
     3721 ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct
     3781 attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg
     3841 gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat ggtcgaggtg
     3901 agccccacgt tctgcttcac tctccccatc tcccccccct ccccaccccc aattttgtat
     3961 ttatttattt tttaattatt ttgtgcagcg atgggggcgg gggggggggg ggggcgcgcg
     4021 ccaggcgggg cggggcgggg cgaggggcgg ggcggggcga ggcggaaagg tgcggcggca
     4081 gccaatcaaa gcggcgcgct ccgaaagttt ccttttatgg cgaggcggcg gcggcggcgg
     4141 ccctataaaa agcgaagcgc gcggcgggcg ggagtcgctg cgcgctgcct tcgccccgtg
     4201 ccccgctccg ccgccgcctc gcgccgcccg ccccggctct gactgaccgc gttactccca
     4261 caggtgagcg ggcgggacgg cccttctcct ccgggctgta attagcgctt ggtttaatga
     4321 cggcttgttt cttttctgtg gctgcgtgaa agccttgagg ggctccggga gcgccggcag
     4381 gaaggaaatg ggcggggagg gccttcgtgc gtcgccgcgc cgccgtcccc ttctccctct
     4441 ccagcctcgg ggctgtccgc ggggggacgg ctgccttcgg gggggacggg gcagggcggg
     4501 gttcggcttc tggcgtgtga ccggcggctc tagagcctct gctaaccatg ttcatgcctt
     4561 cttctttttc ctacagctcc tgggcaacgt gctggttatt gtgctgtctc atcattttgg
     4621 caaagaattg attaattcga gcgaacgcgt gtaccgaagc cgctagcgct accggtcgcc
     4681 accatgcccg ccatgaagat cgagtgccgc atcaccggca ccctgaacgg cgtggagttc
     4741 gagctggtgg gcggcggaga gggcaccccc gagcagggcc gcatgaccaa caagatgaag
     4801 agcaccaaag gcgccctgac cttcagcccc tacctgctga gccacgtgat gggctacggc
     4861 ttctaccact tcggcaccta ccccagcggc tacgagaacc ccttcctgca cgccatcaac
     4921 aacggcggct acaccaacac ccgcatcgag aagtacgagg acggcggcgt gctgcacgtg
     4981 agcttcagct accgctacga ggccggccgc gtgatcggcg acttcaaggt ggtgggcacc
     5041 ggcttccccg aggacagcgt gatcttcacc gacaagatca tccgcagcaa cgccaccgtg
     5101 gagcacctgc accccatggg cgataacgtg ctggtgggca gcttcgcccg caccttcagc
     5161 ctgcgcgacg gcggctacta cagcttcgtg gtggacagcc acatgcactt caagagcgcc
     5221 atccacccca gcatcctgca gaacgggggc cccatgttcg ccttccgccg cgtggaggag
     5281 ctgcacagca acaccgagct gggcatcgtg gagtaccagc acgccttcaa gacccccatc
     5341 gccttcgcca gatctcgagc tcgatgagtt tggacaaacc acaactagaa tgcagtgaaa
     5401 aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg
     5461 caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc agggggaggt
     5521 gtgggaggtt ttttaaagca agtaaaacct ctacaaatgt ggtacttaag agggatcgat
     5581 aaggatctag cttgtggaag gctactcgaa atgtttgacc caagttaaac aatttaaagg
     5641 caatgctacc aaatactaat tgagtggttg gaaacttctg acccccgggg aaggggtgaa
     5701 aaaaaaaaaa ctgaaaggaa ctttc
//
