LOCUS       sequence-309361-        7350 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  SB-transposon with constitutive bi-directional promoter. One side 
            contains tea caffeine synthase 1 (TCS2) from Camellia sinensis; the 
            other side contains GFP and blasticidin resistance genes. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7350)
  TITLE     Methylxanthine Synthesis in Human Cells
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7350)
  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            "Unpublished"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-GB TCS2
FEATURES             Location/Qualifiers
     source          1..7350
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        109..213
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(214..282,283..1074)
                     /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(432..451)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1245..1833
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1734..1753
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1987..2004
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     2378..2397
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     polyA_signal    2396..2451
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     2497..2516
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     CDS             complement(2616..3014)
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAMPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     CDS             complement(3015..3071)
                     /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(join(3081..3791,3792..3794,3795..3797))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MLSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             complement(join(3081..3791,3792..3794,3795..3797))
                     /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(3123..3144)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3472..3491
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3731..3752
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     promoter        4429..5611
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     primer_bind     5559..5579
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     primer_bind     complement(6797..6814)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6803..7027
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
ORIGIN
        1 aattttaaca aaatattaac gcttacaatt tcctgatgcg gtattttctc cttacgcatc
       61 tgtgcggtat ttcacaccgc atcaggtggc acttttcggg gaaatgtgcg cggaacccct
      121 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
      181 taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc
      241 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
      301 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc
      361 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact
      421 tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc
      481 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag
      541 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
      601 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt
      661 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
      721 gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc
      781 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg
      841 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
      901 gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca
      961 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
     1021 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca
     1081 gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     1141 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     1201 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     1261 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     1321 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     1381 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     1441 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     1501 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     1561 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     1621 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     1681 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     1741 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     1801 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     1861 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     1921 gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc
     1981 gagcgcagcg agtcagtgag cgaggaagcg gatccctata cagttgaagt cggaagttta
     2041 catacacctt agccaaatac atttaaactc actttttcac aattcctgac atttaatcct
     2101 agtaaaaatt ccctgtctta ggtcagttag gatcaccact ttattttaag aatgtgaaat
     2161 atcagaataa tagtagagag aatgattcat ttcagctttt atttctttca tcacattccc
     2221 agtgggtcag aagtttacat acactcgact cctctgcaga atgcggcgat gtttcggtaa
     2281 ggggtcccga gtagctagtt catggcagcc agcatatggc atatgttgcc aaactctaaa
     2341 ccaaatactc attctgatgt tttaaatgat ttgccctccc atatgtcctt ccgagtgaga
     2401 gacacaaaaa attccaacac actattgcaa tgaaaataaa tttcctttat tagccagaag
     2461 tcagatgctc aaggggcttc atgatgtccc cataattttt ggcagaggga aaaagatctc
     2521 agtggtattt gtgagccagg gcattggcca caccagccac caccttctga taggcagcct
     2581 gcacctgagg agtgaattca cgacaggcct tcgaattagc cctcccacac ataaccagag
     2641 ggcagcaatt cacgaatccc aactgccgtc ggctgtccat cactgtcctt cactatggct
     2701 ttgatcccag gatgcagatc gagaagcacc tgtcggcacc gtccgcaggg gctcaagatg
     2761 cccctgttct catttccgat cgcgacgata caagtcaggt tgccagctgc cgcagcagca
     2821 gcagtgccca gcaccacgag ttctgcacaa ggtcccccag taaaatgata tacattgaca
     2881 ccagtgaaga tgcggccgtc gctagagaga gctgcgctgg cgacgctgta gtcttcagag
     2941 atggggatgc tgttgattgt agccgttgct ctttcaatga gggtggattc ttcttgagac
     3001 aaaggcatgg ccatggggcc aggattctcc tcgacgtcac cagcctgctt cagcaggctg
     3061 aagttagtag ctccactgcc cttgtacagc tcgtccatgc cgagagtgat cccggcggcg
     3121 gtcacgaact ccagcaggac catgtgatcg cgcttctcgt tggggtcttt gctcagggcg
     3181 gactgggtgc tcaggtagtg gttgtcgggc agcagcacgg ggccgtcgcc gatgggggtg
     3241 ttctgctggt agtggtcggc gagctgcacg ctgccgtcct cgatgttgtg gcggatcttg
     3301 aagttcacct tgatgccgtt cttctgcttg tcggccatga tatagacgtt gtggctgttg
     3361 tagttgtact ccagcttgtg ccccaggatg ttgccgtcct ccttgaagtc gatgcccttc
     3421 agctcgatgc ggttcaccag ggtgtcgccc tcgaacttca cctcggcgcg ggtcttgtag
     3481 ttgccgtcgt ccttgaagaa gatggtgcgc tcctggacgt agccttcggg catggcggac
     3541 ttgaagaagt cgtgctgctt catgtggtcg gggtagcggc tgaagcactg cacgccgtag
     3601 gtcagggtgg tcacgagggt gggccagggc acgggcagct tgccggtggt gcagatgaac
     3661 ttcagggtca gcttgccgta ggtggcatcg ccctcgccct cgccggacac gctgaacttg
     3721 tggccgttta cgtcgccgtc cagctcgacc aggatgggca ccaccccggt gaacagctcc
     3781 tcgcccttgc tcaacatggt ggcctcaggt gcagaggttt ctacagggaa agagagaaag
     3841 gctacagtta gtttgctgtg atgatgctaa gtgccgaggt cagatgtcta tggattgtac
     3901 acggaatccc aacgcaggaa ccaaaaaaca ctcacccaaa aaaatggcgc taaggccgag
     3961 agaaaggcga agttccgtct acggctatat accttgaaac aggggcggca gaagtgacgc
     4021 aacgcggttg cgcggacggc ttcctgtgtt ttcattggtc gaagcgttct gcgcgtgcgc
     4081 agagattccc cagggaaggc gggccctctt ccgaatgtcc aatccggaca ttcttccggt
     4141 tggacctccg cggtcagcgc tgtgcccggc gagagatcac gtggggcgcg gaggcggtgc
     4201 tgctggggca cggccgtcca gcctcggcgg ccatattttt gaggggctgt tcatctcgtt
     4261 cacacgctct gtccgccatg tttgtgagtg gaagcgccat taccccttca agcgactgaa
     4321 ggctgcaggg cctctggtgg cccgcatggg gagaccagac ccgccaggcc cgcctttccg
     4381 cactcagtcc gggcttactt tattttgtga gacagggtct cgcctagagg ctccggtgcc
     4441 cgtcagtggg cagagcgcac atcgcccaca gtccccgaga agttgggggg aggggtcggc
     4501 aattgaaccg gtgcctagag aaggtggcgc ggggtaaact gggaaagtga tgtcgtgtac
     4561 tggctccgcc tttttcccga gggtggggga gaaccgtata taagtgcagt agtcgccgtg
     4621 aacgttcttt ttcgcaacgg gtttgccgcc agaacacagg taagtgccgt gtgtggttcc
     4681 cgcgggcctg gcctctttac gggttatggc ccttgcgtgc cttgaattac ttccacgccc
     4741 ctggctgcag tacgtgattc ttgatcccga gcttcgggtt ggaagtgggt gggagagttc
     4801 gaggccttgc gcttaaggag ccccttcgcc tcgtgcttga gttgaggcct ggcttgggcg
     4861 ctggggccgc cgcgtgcgaa tctggtggca ccttcgcgcc tgtctcgctg ctttcgataa
     4921 gtctctagcc atttaaaatt tttgatgacc tgctgcgacg ctttttttct ggcaagatag
     4981 tcttgtaaat gcgggccaag atctgcacac tggtatttcg gtttttgggg ccgcgggcgg
     5041 cgacggggcc cgtgcgtccc agcgcacatg ttcggcgagg cggggcctgc gagcgcggcc
     5101 accgagaatc ggacgggggt agtctcaagc tggccggcct gctctggtgc ctggcctcgc
     5161 gccgccgtgt atcgccccgc cctgggcggc aaggctggcc cggtcggcac cagttgcgtg
     5221 agcggaaaga tggccgcttc ccggccctgc tgcagggagc tcaaaatgga ggacgcggcg
     5281 ctcgggagag cgggcgggtg agtcacccac acaaaggaaa agggcctttc cgtcctcagc
     5341 cgtcgcttca tgtgactcca cggagtaccg ggcgccgtcc aggcacctcg attagttctc
     5401 gagcttttgg agtacgtcgt ctttaggttg gggggagggg ttttatgcga tggagtttcc
     5461 ccatactgag tgggtggaga ctgaagttag gccagcttgg cacttgatgt aattctcctt
     5521 ggaatttgcc ctttttgagt ttggatcttg gttcattctc aagcctcaga cagtggttca
     5581 aagttttttt cttccattta aggtgtcgtg aaaactaccc caagctggcc tctgaggcca
     5641 ccatgaagga agtgaaagag gccctcttca tgaacaaagg cgagggcgag agcagctacg
     5701 cccagaatag cagctttacc cagaccgtga ccagcatgac catgcctgtg ctggaaaacg
     5761 ccgtggaaac cctgttcagc aaggacttcc atctgctgca ggccctgaat gccgtggatc
     5821 tgggatgtgc tgccggacct accaccttca ccgtgatcag caccatcaag cggatgatgg
     5881 aaaagaagtg ccgcgagctg aactgtcaga ccctggaact gcaggtctac ctgaacgacc
     5941 tgcctggcaa cgacttcaac accctgttta agggcctgcc tagcaaggtc gtgggcaaca
     6001 agtgtgaaga ggtgtcctgc tacgtcgtgg gagtgcctgg ctcttttcac ggcagactgt
     6061 tccccagaaa cagcctgcac ctggtgcaca gctgttactc cgtgcactgg ctgacacagg
     6121 ccccaaaggg cctgacctcc aaagaaggac tggccctgaa caagggcaag atctacatca
     6181 gcaagacaag ccctccagtc gtgcgcgagg cctacctgtc tcagttccac gaggacttca
     6241 ccatgtttct gaacagccgc agccaagagg tggtgcccaa tggctgtatg gtcctgatcc
     6301 tgagaggccg gctgagcagc gatcctagcg atatgggcag ctgcttcacc tgggaactgc
     6361 tggccgtggc cattgccgaa ctggtttctc agggactgat cgacgaggac aagctggaca
     6421 ccttcaacgt gcccagctac ttccccagcc tggaagaagt gaaggacatc gtcgagcgga
     6481 acggcagctt caccatcgac cacatggaag gcttcgagct ggacagcccc gagatgcaag
     6541 agaacgacaa atgggtccga ggcgagaagt tcgccacagt ggccagagcc tttaccgagc
     6601 ctatcatcag caatcagttc ggccacgaga tcatggataa gctgtacgag aagtttaccc
     6661 acatcgtggt gtccgacttc gaggccaaga ttcccaagat caccagcatc atcctggtgc
     6721 tgtccaagat cgtgggctaa ttcgaaggcc tgtcaggcca agcttgggga tcaattctct
     6781 agagctcgct gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc
     6841 tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat
     6901 gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg
     6961 caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc
     7021 tctatggctt ctgaggcgga aagaaccagc tgggcctact agatagcgga ccccttaccg
     7081 aaacatcgcc gcattctgca gaggagtcga acttagtgta tgtaaacttc tgacccactg
     7141 gaattgtgat acagtgaatt ataagtgaaa taatctgtct gtaaacaatt gttggaaaaa
     7201 tgacttgtgt catgcacaaa gtagatgtcc taactgactt gccaaaacta ttgtttgtta
     7261 acaagaaatt tgtggagtag ttgaaaaacg agttttaatg actccaactt aagtgtatgt
     7321 aaacttccga cttcaactgt atagggatcc
//
