LOCUS       sequence-133642-        5036 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Sleeping Beauty (SB) transposon with G418 resistance.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5036)
  AUTHORS   Chicaybam L, Barcelos C, Peixoto B, Carneiro M, Limia CG, Redondo P,
            Lira C, Paraguassu-Braga F, Vasconcelos ZF, Barros L, Bonamino MH
  TITLE     An Efficient Electroporation Protocol for the Genetic Modification 
            of Mammalian Cells.
  JOURNAL   Front Bioeng Biotechnol. 2017 Jan 23;4:99. doi: 
            10.3389/fbioe.2016.00099. eCollection 2016.
  PUBMED    28168187
REFERENCE   2  (bases 1 to 5036)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Front 
            Bioeng Biotechnol."; date: "2017-01-23"; volume: "4:99. doi"; pages:
            " 10.3389/fbioe.2016.00099. eCollection 2016"
COMMENT     Sequence Label: pT3-Neo
FEATURES             Location/Qualifiers
     source          1..5036
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(406..422)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(406..422)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(419..441)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    430..446
                     /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(join(454..460,461..478,479..484))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    499..520
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(637..654)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(808..1396)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(888..907)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1567..2358,2359..2427))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     2190..2209
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2428..2532)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2600..2618
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2656..2678)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2778..2797
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2991..3013
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3006..3022
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3404..3424)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     regulatory      3900..3909
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(4735..4753)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(4736..4753)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(4773..4790)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4779..5003
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
ORIGIN
        1 gatccccttg aaatacatcc acaggtacac ctccaattga ctcaaatgat gtcaattagt
       61 ctatcagaag cttctaaagc catgacatca ttttctggaa ttttccaagc tgtttaaagg
      121 cacagtcaac ttagtgtatg taaacttctg acccactgga attgtgatac agtgaattat
      181 aagtgaaata atctgtctgt aaacaattgt tggaaaaatg acttgtgtca tgcacaaagt
      241 agatgtccta actgacttgc caaaactatt gtttgttaac aagaaatttg tggagtagtt
      301 gaaaaacgag ttttaatgac tccaacttaa gtgtatgtaa acttccgact tcaactgtat
      361 aggtctagag tcgacctgca ggcatgcaag cttggcgtaa tcatggtcat agctgtttcc
      421 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
      481 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
      541 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
      601 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
      661 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
      721 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
      781 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
      841 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
      901 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
      961 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct caatgctcac gctgtaggta
     1021 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     1081 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     1141 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     1201 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg
     1261 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     1321 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     1381 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     1441 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     1501 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     1561 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     1621 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     1681 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     1741 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     1801 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     1861 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     1921 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     1981 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     2041 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     2101 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     2161 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     2221 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     2281 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     2341 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     2401 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     2461 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     2521 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat
     2581 catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg
     2641 tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta
     2701 agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg
     2761 gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc atatgcggtg
     2821 tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggcgccatt cgccattcag
     2881 gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctggc
     2941 gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt cccagtcacg
     3001 acgttgtaaa acgacggcca gtgaattcga gctcggtacc ctacagttga agtcggaagt
     3061 ttacatacac ttaagttgga gtcattaaaa ctcgtttttc aactactcca caaatttctt
     3121 gttaacaaac aatagttttg gcaagtcagt taggacatct actttgtgca tgacacaagt
     3181 catttttcca acaattgttt acagacagat tatttcactt ataattcact gtatcacaat
     3241 tccagtgggt cagaagttta catacactaa gttgactgtg cctttaaaca gcttggaaaa
     3301 ttccagaaaa tgatgtcatg gctttagaag cttctgatag actaattgac atcatttgag
     3361 tcaattggag gtgtacctgt ggatgtattt caaggaattc tgtggaatgt gtgtcagtta
     3421 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatatcgat
     3481 actagtttaa ttaagatctc gagggaatga aagaccccac ctgtaggttt ggcaagctag
     3541 cttaagtaac gccattttgc aaggcatgga aaatacataa ctgagaatag agaagttcag
     3601 atcaaggtta ggaacagaga gacagcagaa tatgggccaa acaggatatc tgtggtaagc
     3661 agttcctgcc ccggctcagg gccaagaaca gatggtcccc agatgcggtc ccgccctcag
     3721 cagtttctag agaaccatca gatgtttcca gggtgcccca aggacctgaa atgaccctgt
     3781 gccttatttg aactaaccaa tcagttcgct tctcgcttct gttcgcgcgc ttctgctccc
     3841 cgagctcaat aaaagagccc acaacccctc actcggcgcg ccagtccgga tccaccggtg
     3901 ccaccatggc cgaacaggat ggactgcacg caggctcccc cgccgcttgg gtggaaagac
     3961 tgtttggata cgactgggct cagcagacaa tcggatgctc tgacgcagcc gtgttccggc
     4021 tgagtgctca ggggcgcccc gtgctgtttg tcaagacaga cctgtcagga gcactgaacg
     4081 agctgcagga tgaagcagcc cgcctgagct ggctggctac cacaggagtg ccttgtgctg
     4141 cagtcctgga cgtggtcact gaggcaggcc gagattggct gctgctgggg gaagtgccag
     4201 gacaggacct gctgagctcc cacctggcac cagctgagaa ggtctccatc atggcagatg
     4261 ccatgcggag actgcatact ctggaccccg ccacctgccc tttcgatcac caggctaaac
     4321 atcgcattga gagggctcgc acccgaatgg aagcaggcct ggtggaccag gacgatctgg
     4381 atgaggaaca ccaggggctg gctcctgcag agctgtttgc acggctgaaa gccagaatgc
     4441 cagacggaga agatctggtg gtcacccatg gcgacgcttg cctgcccaac atcatggtgg
     4501 agaatggcag attcagtggg tttattgatt gtggacgact gggcgtcgcc gacagatacc
     4561 aggatatcgc cctggctaca agagacattg cagaggaact gggcggggaa tgggccgata
     4621 ggttcctggt gctgtatggc atcgctgccc ctgattcaca gaggattgcc ttttacagac
     4681 tgctggatga gttcttttga gcggccgctc gagcatgcat ctagagggcc ctattctata
     4741 gtgtcaccta aatgctagag ctcgctgatc agcctcgact gtgccttcta gttgccagcc
     4801 atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt
     4861 cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct
     4921 ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc
     4981 tggggatgcg gtgggctcta tggcttctga ggcggaaaga accagctgca ttaatg
//
