LOCUS       sequence-97532-e        7066 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  SB-transposon with constitutive bi-directional promoter, one side: 
            SfiI cloning site for GOI (contains filler DNA), other side: GFP and
            hygromycin resistance gene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7066)
  AUTHORS   Kowarz E, Loescher D, Marschalek R
  TITLE     Optimized Sleeping Beauty transposons rapidly generate stable 
            transgenic cell lines.
  JOURNAL   Biotechnol J. 2015 Feb 4. doi: 10.1002/biot.201400821.
  PUBMED    25650551
REFERENCE   2  (bases 1 to 7066)
  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: "Biotechnol 
            J. 2015 Feb 4. doi: 10.1002/biot.201400821."
COMMENT     Sequence Label: pSBbi-GH
FEATURES             Location/Qualifiers
     source          1..7066
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    324..548
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     537..554
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     regulatory      794..803
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(829..2011)
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     primer_bind     complement(861..881)
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             join(2643..2645,2646..2648,2649..3359)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MLSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(2643..2645,2646..2648,2649..3359)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MLSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2688..2709)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2949..2968)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3296..3317
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             3369..3425
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             3426..4451
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             3426..4451
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     primer_bind     complement(4551..4570)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4616..4671
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4670..4689)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5063..5080)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5234..5822)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5314..5333)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5993..6784,6785..6853))
                     /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     6616..6635
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6854..6958)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ggatccctat acagttgaag tcggaagttt acatacactt aagttggagt cattaaaact
       61 cgtttttcaa ctactccaca aatttcttgt taacaaacaa tagttttggc aagtcagtta
      121 ggacatctac tttgtgcatg acacaagtca tttttccaac aattgtttac agacagatta
      181 tttcacttat aattcactgt atcacaattc cagtgggtca gaagtttaca tacactaagt
      241 tcgactcctc tgcagaatgc ggcgatgttt cggtaagggg tccgctatct agtaggccca
      301 gctggttctt tccgcctcag aagccataga gcccaccgca tccccagcat gcctgctatt
      361 gtcttcccaa tcctccccct tgctgtcctg ccccacccca ccccccagaa tagaatgaca
      421 cctactcaga caatgcgatg caatttcctc attttattag gaaaggacag tgggagtggc
      481 accttccagg gtcaaggaag gcacggggga ggggcaaaca acagatggct ggcaactaga
      541 aggcacagtc gaggctgatc agcgagctct agagaattga tccccaagct tggcctgaca
      601 ggccttcgaa ttagagagaa ggaaagacgc aaccaggaag cccaccaaga gaaagaggca
      661 tttcctgaaa agattcccct ttttggagag ccctacaagg tatttactga acactagaca
      721 ttgaagtcct gatttatcac aatgttgaac cctatgatga aacaattcag tataattgag
      781 ttcgaaactc gagccatggt ggcctcagag gccagcttgg ggtagttttc acgacacctt
      841 aaatggaaga aaaaaacttt gaaccactgt ctgaggcttg agaatgaacc aagatccaaa
      901 ctcaaaaagg gcaaattcca aggagaatta catcaagtgc caagctggcc taacttcagt
      961 ctccacccac tcagtatggg gaaactccat cgcataaaac ccctcccccc aacctaaaga
     1021 cgacgtactc caaaagctcg agaactaatc gaggtgcctg gacggcgccc ggtactccgt
     1081 ggagtcacat gaagcgacgg ctgaggacgg aaaggccctt ttcctttgtg tgggtgactc
     1141 acccgcccgc tctcccgagc gccgcgtcct ccattttgag ctccctgcag cagggccggg
     1201 aagcggccat ctttccgctc acgcaactgg tgccgaccgg gccagccttg ccgcccaggg
     1261 cggggcgata cacggcggcg cgaggccagg caccagagca ggccggccag cttgagacta
     1321 cccccgtccg attctcggtg gccgcgctcg caggccccgc ctcgccgaac atgtgcgctg
     1381 ggacgcacgg gccccgtcgc cgcccgcggc cccaaaaacc gaaataccag tgtgcagatc
     1441 ttggcccgca tttacaagac tatcttgcca gaaaaaaagc gtcgcagcag gtcatcaaaa
     1501 attttaaatg gctagagact tatcgaaagc agcgagacag gcgcgaaggt gccaccagat
     1561 tcgcacgcgg cggccccagc gcccaggcca agcctcaact caagcacgag gcgaaggggc
     1621 tccttaagcg caaggcctcg aactctccca cccacttcca acccgaagct cgggatcaag
     1681 aatcacgtac tgcagccagg ggcgtggaag taattcaagg cacgcaaggg ccataacccg
     1741 taaagaggcc aggcccgcgg gaaccacaca cggcacttac ctgtgttctg gcggcaaacc
     1801 cgttgcgaaa aagaacgttc acggcgacta ctgcacttat atacggttct cccccaccct
     1861 cgggaaaaag gcggagccag tacacgacat cactttccca gtttaccccg cgccaccttc
     1921 tctaggcacc ggttcaattg ccgacccctc cccccaactt ctcggggact gtgggcgatg
     1981 tgcgctctgc ccactgacgg gcaccggagc ctctaggcga gaccctgtct cacaaaataa
     2041 agtaagcccg gactgagtgc ggaaaggcgg gcctggcggg tctggtctcc ccatgcgggc
     2101 caccagaggc cctgcagcct tcagtcgctt gaaggggtaa tggcgcttcc actcacaaac
     2161 atggcggaca gagcgtgtga acgagatgaa cagcccctca aaaatatggc cgccgaggct
     2221 ggacggccgt gccccagcag caccgcctcc gcgccccacg tgatctctcg ccgggcacag
     2281 cgctgaccgc ggaggtccaa ccggaagaat gtccggattg gacattcgga agagggcccg
     2341 ccttccctgg ggaatctctg cgcacgcgca gaacgcttcg accaatgaaa acacaggaag
     2401 ccgtccgcgc aaccgcgttg cgtcacttct gccgcccctg tttcaaggta tatagccgta
     2461 gacggaactt cgcctttctc tcggccttag cgccattttt ttgggtgagt gttttttggt
     2521 tcctgcgttg ggattccgtg tacaatccat agacatctga cctcggcact tagcatcatc
     2581 acagcaaact aactgtagcc tttctctctt tccctgtaga aacctctgca cctgaggcca
     2641 ccatgttgag caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg
     2701 acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct
     2761 acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca
     2821 ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc gaccacatga
     2881 agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct
     2941 tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc
     3001 tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc
     3061 acaagctgga gtacaactac aacagccaca acgtctatat catggccgac aagcagaaga
     3121 acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc gtgcagctcg
     3181 ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc
     3241 actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc gatcacatgg
     3301 tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagg
     3361 gcagtggagc tactaacttc agcctgctga agcaggctgg tgacgtcgag gagaatcctg
     3421 gccccatgaa aaagcctgaa ctcaccgcga cgtctgtcga gaagtttctg atcgaaaagt
     3481 tcgacagcgt ctccgacctg atgcagctct cggagggcga agaatctcgt gctttcagct
     3541 tcgatgtagg agggcgtgga tatgtcctgc gggtaaatag ctgcgccgat ggtttctaca
     3601 aagatcgtta tgtttatcgg cactttgcat cggccgcgct cccgattccg gaagtgcttg
     3661 acattgggga attcagcgag agcctgacct attgcatctc ccgccgtgca cagggtgtca
     3721 cgttgcaaga cctgcctgaa accgaactgc ccgctgttct gcagccggtc gcggaggcca
     3781 tggatgcgat cgctgcggcc gatcttagcc agacgagcgg gttcggccca ttcggaccgc
     3841 aaggaatcgg tcaatacact acatggcgtg atttcatatg cgcgattgct gatccccatg
     3901 tgtatcactg gcaaactgtg atggacgaca ccgtcagtgc gtccgtcgcg caggctctcg
     3961 atgagctgat gctttgggcc gaggactgcc ccgaagtccg gcacctcgtg cacgcggatt
     4021 tcggctccaa caatgtcctg acggacaatg gccgcataac agcggtcatt gactggagcg
     4081 aggcgatgtt cggggattcc caatacgagg tcgccaacat cttcttctgg aggccgtggt
     4141 tggcttgtat ggagcagcag acgcgctact tcgagcggag gcatccggag cttgcaggat
     4201 cgccgcggct ccgggcgtat atgctccgca ttggtcttga ccaactctat cagagcttgg
     4261 ttgacggcaa tttcgatgat gcagcttggg cgcagggtcg atgcgacgca atcgtccgat
     4321 ccggagccgg gactgtcggg cgtacacaaa tcgcccgcag aagcgcggcc gtctggaccg
     4381 atggctgtgt agaagtactc gccgatagtg gaaaccgacg ccccagcact cgtccgaggg
     4441 caaaggaata gttcgaaggc ctgtcgtgaa ttcactcctc aggtgcaggc tgcctatcag
     4501 aaggtggtgg ctggtgtggc caatgccctg gctcacaaat accactgaga tctttttccc
     4561 tctgccaaaa attatgggga catcatgaag ccccttgagc atctgacttc tggctaataa
     4621 aggaaattta ttttcattgc aatagtgtgt tggaattttt tgtgtctctc actcggaagg
     4681 acatatggga gggcaaatca tttaaaacat cagaatgagt atttggttta gagtttggca
     4741 acatatgcca tatgctggct gccatgaact agctactcgg gaccccttac cgaaacatcg
     4801 ccgcattctg cagaggagtc gagtgtatgt aaacttctga cccactggga atgtgatgaa
     4861 agaaataaaa gctgaaatga atcattctct ctactattat tctgatattt cacattctta
     4921 aaataaagtg gtgatcctaa ctgacctaag acagggaatt tttactagga ttaaatgtca
     4981 ggaattgtga aaaagtgagt ttaaatgtat ttggctaagg tgtatgtaaa cttccgactt
     5041 caactgtata gggatccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     5101 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     5161 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     5221 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     5281 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     5341 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     5401 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     5461 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     5521 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     5581 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     5641 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     5701 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     5761 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     5821 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     5881 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     5941 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     6001 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     6061 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     6121 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     6181 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     6241 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     6301 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     6361 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     6421 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     6481 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     6541 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     6601 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     6661 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     6721 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     6781 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     6841 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     6901 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     6961 catttccccg aaaagtgcca cctgatgcgg tgtgaaatac cgcacagatg cgtaaggaga
     7021 aaataccgca tcaggaaatt gtaagcgtta atattttgtt aaaatt
//
