LOCUS       sequence-309356-        8250 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  SB-transposon with constitutive bi-directional promoter. One side 
            contains N-GST tea caffeine synthase 1 (TCS1) from Camellia 
            sinensis; the other side contains RFP and puromycin resistance 
            genes..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8250)
  TITLE     Methylxanthine Synthesis in Human Cells
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8250)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pSBBi-RP GST-TCS1
FEATURES             Location/Qualifiers
     source          1..8250
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..105
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(106..174,175..966)
                     /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     complement(324..343)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1137..1725
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1626..1645
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1879..1896
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     2270..2289
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     polyA_signal    2288..2343
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     2389..2408
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     CDS             complement(2508..3107)
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(2591..2611)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     3088..3107
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     CDS             complement(3108..3164)
                     /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             complement(3174..3875)
                     /codon_start=1
                     /product="dimeric variant of DsRed fluorescent protein 
                     (Shaner et al., 2004)"
                     /label=dTomato
                     /note="mammalian codon-optimized"
                     /translation="MLSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     LVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH
                     HLFLYGMDELYK"
     primer_bind     complement(3257..3280)
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     primer_bind     complement(3455..3474)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     3724..3742
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     promoter        4507..5689
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     primer_bind     5637..5657
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             5721..6374
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     6332..6354
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             6393..6413
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(7589..7606)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    7595..7819
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
ORIGIN
        1 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
       61 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
      121 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
      181 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
      241 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
      301 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg
      361 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
      421 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
      481 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
      541 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
      601 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca
      661 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
      721 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
      781 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
      841 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
      901 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
      961 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
     1021 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     1081 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     1141 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     1201 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     1261 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     1321 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     1381 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     1441 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     1501 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     1561 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     1621 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     1681 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     1741 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     1801 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
     1861 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggatcccta tacagttgaa
     1921 gtcggaagtt tacatacacc ttagccaaat acatttaaac tcactttttc acaattcctg
     1981 acatttaatc ctagtaaaaa ttccctgtct taggtcagtt aggatcacca ctttatttta
     2041 agaatgtgaa atatcagaat aatagtagag agaatgattc atttcagctt ttatttcttt
     2101 catcacattc ccagtgggtc agaagtttac atacactcga ctcctctgca gaatgcggcg
     2161 atgtttcggt aaggggtccc gagtagctag ttcatggcag ccagcatatg gcatatgttg
     2221 ccaaactcta aaccaaatac tcattctgat gttttaaatg atttgccctc ccatatgtcc
     2281 ttccgagtga gagacacaaa aaattccaac acactattgc aatgaaaata aatttccttt
     2341 attagccaga agtcagatgc tcaaggggct tcatgatgtc cccataattt ttggcagagg
     2401 gaaaaagatc tcagtggtat ttgtgagcca gggcattggc cacaccagcc accaccttct
     2461 gataggcagc ctgcacctga ggagtgaatt cacgacaggc cttcgaatca ggcaccgggc
     2521 ttgcgggtca tgcaccaggt gcgcggtcct tcgggcacct cgacgtcggc ggtgacggtg
     2581 aagccgagcc gctcgtagaa ggggaggttg cggggcgcgg aggtctccag gaaggcgggc
     2641 accccggcgc gctcggccgc ctccactccg gggagcacga cggcgctgcc cagacccttg
     2701 ccctggtggt cgggcgagac gccgacggtg gccaggaacc acgcgggctc cttgggccgg
     2761 tgcggcgcca ggaggccttc catctgttgc tgcgcggcca gccgggaacc gctcaactcg
     2821 gccatgcgcg ggccgatctc ggcgaacacc gcccccgctt cgacgctctc cggcgtggtc
     2881 cagaccgcca ccgcggcgcc gtcgtccgcg acccacacct tgccgatgtc gagcccgacg
     2941 cgcgtgagga agagttcttg cagctcggtg acccgctcga tgtggcggtc cggatcgacg
     3001 gtgtggcgcg tggcggggta gtcggcgaac gcggcggcga gggtgcgtac ggccctgggg
     3061 acgtcgtcgc gggtggcgag gcgcaccgtg ggcttgtact cggtcatggg gccaggattc
     3121 tcctcgacgt caccagcctg cttcagcagg ctgaagttag tagctccact gcccttgtac
     3181 agctcgtcca tgccgtacag gaacaggtgg tggcggccct cggagcgctc gtactgttcc
     3241 acgatggtgt agtcctcgtt gtgggaggtg atgtccagct tggtgtccac gtagtagtag
     3301 ccgggcagtt gcacgggctt cttggccatg tagatggtct tgaactccac caggtagtgg
     3361 ccgccgtcct tcagcttcag ggcctggtgg atctcgccct tcagcacgcc gtcgcggggg
     3421 tacaggcgct cggtggaggc ctcccagccc attgtcttct tctgcattac ggggccgtcg
     3481 ggggggaagt tggtgccgcg catcttcacc ttgtagatca gcgtgccgtc ctgcagggag
     3541 gagtcctggg tcacggtcac cagaccgccg tcctcgaagt tcatcacgcg ctcccacttg
     3601 aagccctcgg ggaaggacag cttcttgtaa tcggggatgt cggcggggtg cttcacgtac
     3661 gccttggagc cgtacatgaa ctggggggac aggatgtccc aggcgaaggg cagggggccg
     3721 cccttggtca ccttcagctt ggcggtctgg gtgccctcgt aggggcggcc ctcgccctcg
     3781 ccctcgatct cgaactcatg gccgttcatg gagccctcca tgcgcacctt gaagcgcatg
     3841 aactctttga tgacctcctc gcccttgctc aacatggtgg cctcaggtgc agaggtttct
     3901 acagggaaag agagaaaggc tacagttagt ttgctgtgat gatgctaagt gccgaggtca
     3961 gatgtctatg gattgtacac ggaatcccaa cgcaggaacc aaaaaacact cacccaaaaa
     4021 aatggcgcta aggccgagag aaaggcgaag ttccgtctac ggctatatac cttgaaacag
     4081 gggcggcaga agtgacgcaa cgcggttgcg cggacggctt cctgtgtttt cattggtcga
     4141 agcgttctgc gcgtgcgcag agattcccca gggaaggcgg gccctcttcc gaatgtccaa
     4201 tccggacatt cttccggttg gacctccgcg gtcagcgctg tgcccggcga gagatcacgt
     4261 ggggcgcgga ggcggtgctg ctggggcacg gccgtccagc ctcggcggcc atatttttga
     4321 ggggctgttc atctcgttca cacgctctgt ccgccatgtt tgtgagtgga agcgccatta
     4381 ccccttcaag cgactgaagg ctgcagggcc tctggtggcc cgcatgggga gaccagaccc
     4441 gccaggcccg cctttccgca ctcagtccgg gcttacttta ttttgtgaga cagggtctcg
     4501 cctagaggct ccggtgcccg tcagtgggca gagcgcacat cgcccacagt ccccgagaag
     4561 ttggggggag gggtcggcaa ttgaaccggt gcctagagaa ggtggcgcgg ggtaaactgg
     4621 gaaagtgatg tcgtgtactg gctccgcctt tttcccgagg gtgggggaga accgtatata
     4681 agtgcagtag tcgccgtgaa cgttcttttt cgcaacgggt ttgccgccag aacacaggta
     4741 agtgccgtgt gtggttcccg cgggcctggc ctctttacgg gttatggccc ttgcgtgcct
     4801 tgaattactt ccacgcccct ggctgcagta cgtgattctt gatcccgagc ttcgggttgg
     4861 aagtgggtgg gagagttcga ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt
     4921 tgaggcctgg cttgggcgct ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg
     4981 tctcgctgct ttcgataagt ctctagccat ttaaaatttt tgatgacctg ctgcgacgct
     5041 ttttttctgg caagatagtc ttgtaaatgc gggccaagat ctgcacactg gtatttcggt
     5101 ttttggggcc gcgggcggcg acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg
     5161 gggcctgcga gcgcggccac cgagaatcgg acgggggtag tctcaagctg gccggcctgc
     5221 tctggtgcct ggcctcgcgc cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg
     5281 gtcggcacca gttgcgtgag cggaaagatg gccgcttccc ggccctgctg cagggagctc
     5341 aaaatggagg acgcggcgct cgggagagcg ggcgggtgag tcacccacac aaaggaaaag
     5401 ggcctttccg tcctcagccg tcgcttcatg tgactccacg gagtaccggg cgccgtccag
     5461 gcacctcgat tagttctcga gcttttggag tacgtcgtct ttaggttggg gggaggggtt
     5521 ttatgcgatg gagtttcccc atactgagtg ggtggagact gaagttaggc cagcttggca
     5581 cttgatgtaa ttctccttgg aatttgccct ttttgagttt ggatcttggt tcattctcaa
     5641 gcctcagaca gtggttcaaa gtttttttct tccatttaag gtgtcgtgaa aactacccca
     5701 agctggcctc tgaggccacc atgtccccta tactaggtta ttggaaaatt aagggccttg
     5761 tgcaacccac tcgacttctt ttggaatatc ttgaagaaaa atatgaagag catttgtatg
     5821 agcgcgatga aggtgataaa tggcgaaaca aaaagtttga attgggtttg gagtttccca
     5881 atcttcctta ttatattgat ggtgatgtta aattaacaca gtctatggcc atcatacgtt
     5941 atatagctga caagcacaac atgttgggtg gttgtccaaa agagcgtgca gagatttcaa
     6001 tgcttgaagg agcggttttg gatattagat acggtgtttc gagaattgca tatagtaaag
     6061 actttgaaac tctcaaagtt gattttctta gcaagctacc tgaaatgctg aaaatgttcg
     6121 aagatcgttt atgtcataaa acatatttaa atggtgatca tgtaacccat cctgacttca
     6181 tgttgtatga cgctcttgat gttgttttat acatggaccc aatgtgcctg gatgcgttcc
     6241 caaaattagt ttgttttaaa aaacgtattg aagctatccc acaaattgat aagtacttga
     6301 aatccagcaa gtatatagca tggcctttgc agggctggca agccacgttt ggtggtggcg
     6361 accatcctcc aaaatcggat agcagcggca gagaaaactt gtatttccag ggcagagacc
     6421 ctgaaagatc cgagctagct actgcgggga aggtgaacga agtgttgttc atgaacagag
     6481 gagaaggaga aagtagttat gcacaaaact cttctttcac gcaacaagtg gcctcaatgg
     6541 cacagccagc gctagaaaat gcagttgaaa ctctcttctc cagagatttc caccttcaag
     6601 ctcttaacgc agcggacttg ggttgtgcag cgggtccaaa cacattcgca gtgatttcta
     6661 cgatcaagag aatgatggaa aagaaatgca gggaattgaa ttgccaaaca ctggaacttc
     6721 aggtttactt gaatgatctt tttggaaatg atttcaatac cctcttcaaa ggcctgtcgt
     6781 ctgaggttat tggtaacaaa tgtgaggaag ttccgtgtta tgtgatggga gtaccggggt
     6841 ctttccatgg ccggcttttt cctcgtaaca gcttacattt agttcattcc tcttacagtg
     6901 ttcattggct tactcaggca ccaaaaggac tcacaagcag agaaggcttg gcattaaaca
     6961 aggggaagat ttacatatca aagacaagcc ctcctgttgt aagagaagcc tacttatctc
     7021 aatttcatga agatttcaca atgtttctca atgctagatc ccaagaggtg gttccaaatg
     7081 gttgtatggt gttgatactt cgtggtaggc aatgttctga tccttcagac atgcagagct
     7141 gctttacttg ggaactatta gctatggcca ttgctgaatt ggtttcacag ggattgatag
     7201 atgaagataa attagacacc ttcaatatac ccagctattt tgcatcactt gaggaagtga
     7261 aagatatagt ggagagggac ggatcattca caattgatca tatagagggg tttgatcttg
     7321 atagcgtaga aatgcaggag aatgataaat gggttagagg ggaaaagttt accaaggttg
     7381 tcagggcctt cacagagcct ataatttcaa accagtttgg acctgaaatc atggacaaac
     7441 tatatgacaa attcactcac attgtagttt cagatttgga agcaaagcta ccgaagacca
     7501 caagtatcat cctagtgctt tccaagattg atggataggg cctgtcaggc caagcttggg
     7561 gatcaattct ctagagctcg ctgatcagcc tcgactgtgc cttctagttg ccagccatct
     7621 gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt
     7681 tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg
     7741 ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg
     7801 gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gctgggccta ctagatagcg
     7861 gaccccttac cgaaacatcg ccgcattctg cagaggagtc gaacttagtg tatgtaaact
     7921 tctgacccac tggaattgtg atacagtgaa ttataagtga aataatctgt ctgtaaacaa
     7981 ttgttggaaa aatgacttgt gtcatgcaca aagtagatgt cctaactgac ttgccaaaac
     8041 tattgtttgt taacaagaaa tttgtggagt agttgaaaaa cgagttttaa tgactccaac
     8101 ttaagtgtat gtaaacttcc gacttcaact gtatagggat ccaattttaa caaaatatta
     8161 acgcttacaa tttcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc
     8221 gcatcaggtg gcacttttcg gggaaatgtg
//
