LOCUS       sequence-309396-        5679 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  SB-transposon with constitutive bleomycin resistance and 
            Gal4-inducible coffee caffeine synthase 1 (CCS1)..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5679)
  TITLE     Methylxanthine Synthesis in Human Cells
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5679)
  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            "Unpublished"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: pSB-Bleo Gal4 CCS1
FEATURES             Location/Qualifiers
     source          1..5679
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(295..1086,1087..1155))
                     /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     918..937
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1156..1260)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1862..1886
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      1977..1986
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    3182..3303
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3193..3212)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3247..3266
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     regulatory      4057..4066
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4063..4437
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     primer_bind     complement(4492..4509)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     primer_bind     complement(5044..5061)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(5215..5679,1..124))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5295..5314)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
       61 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
      121 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaaca aaaactcacg
      181 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
      241 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
      301 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
      361 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
      421 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
      481 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
      541 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
      601 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
      661 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
      721 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
      781 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
      841 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
      901 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
      961 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     1021 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     1081 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     1141 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     1201 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     1261 cacatttccc cgaaaagtgc cacctgatgc ggtgtgaaat accgcacaga tgcgtaagga
     1321 gaaaataccg catcaggaaa ttgtaagcgt taatattttg ttaaaattgg atccctatac
     1381 agttgaagtc ggaagtttac atacacttaa gttggagtca ttaaaactcg tttttcaact
     1441 actccacaaa tttcttgtta acaaacaata gttttggcaa gtcagttagg acatctactt
     1501 tgtgcatgac acaagtcatt tttccaacaa ttgtttacag acagattatt tcacttataa
     1561 ttcactgtat cacaattcca gtgggtcaga agtttacata cactaagttc gactcctctg
     1621 cagaatgcgg cgatgtttcg gtaaggggtc cgctatctag tgagctctta cgcgtgctag
     1681 ccttgatatc gaattcggag cactgtcctc cgaacgtcgg agcactgtcc tccgaacgtc
     1741 ggagcactgt cctccgaacg tcggagcact gtcctccgaa cggagcatgt cctccgaacg
     1801 tcggagcact gtcctccgaa cgactagtta ggcgtgtacg gtgggaggcc tatataagca
     1861 gagctcgttt agtgaaccgt cagatcgcct ggagacgcca tccacgctgt tttgacctcc
     1921 atagaagaca ccgggaccga tccagcctct cgacattcgt tggatcggcc tctgaggcca
     1981 ccatggaact gcaagaggtg ctgcacatga atggcggcga gggcgataca agctacgcca
     2041 agaacagcag ctacaacctg ttcctgatca gagtgaagcc cgtgctggaa cagtgcatcc
     2101 aagagctgct gagagccaac ctgcctaaca tcaacaagtg cttcaaagtg ggcgacctgg
     2161 gctgtgccag cggacctaat acctttagca ccgtgcggga catcgtgcag agcatcgaca
     2221 aagtgggaca agagaagaag aacgagctgg aacggcctac catccagatc tttctgaacg
     2281 acctgttcca gaacgacttc aacagcgtgt tcaagctgct gcccagcttc taccggaacc
     2341 tggaaaaaga gaacggccgg aagatcggct cctgcctgat tggagctatg cccggctcct
     2401 tctacagcag actgttcccc gaggaatcca tgcactttct gcacagctgc tactgcctgc
     2461 actggctgtc tcaggtgcca tctggcctgg ttacagagct gggcatctct gccaacaagg
     2521 gctgcatcta cagctccaag gccagcgggc ctcctatcaa gaaggcctac ctggaccagt
     2581 tcaccaagga cttcaccacc tttctgcgga tccacagcga ggaactgatc agcagaggcc
     2641 ggatgctgct gaccttcatc tgcaaagagg acgagttcga tcaccccaac agcatggacc
     2701 tgctggaaat gagcatcaac gacctggtca tcgagggcca cctggaagag gaaaagctgg
     2761 acagcttcaa cgtgcccatc tacgccccta gcaccgagga agtgaagcgg atcgttgagg
     2821 aagagggcag cttcgagatc ctgtacctgg aaacctttta cgccccttac gacgccggct
     2881 tcagcatcga cgatgactac cagggcagat ctcacagccc tgtgtcctgt gatgagcacg
     2941 ccagagctgc tcacgtggca tctgttgtgc ggagcatcta cgagcctatc ctggcctctc
     3001 acttcggcga ggccattctg cctgatctga gccaccggat cgccaagaat gccgccaagg
     3061 tgctgagaag cggcaagggc ttttacgaca gcgtgatcat cagcctggcc aagaagcccg
     3121 agaaggccga tatgtagggt aactattcga aggcctgtca ggccgcttcg agcagacatg
     3181 ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt
     3241 atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa
     3301 gttaacaaca acaattgcat tcattttatg tttcaggttc agggggaggt gtgggaggtt
     3361 ttttaaagca agtaaaacct ctacaaatgt ggtaaaatcg ataaggatcc gtcgaccggt
     3421 acctaggcga gaccctgtca caaaataaag taagcccgga ctgagtgcgg aaaggcgggc
     3481 ctggcgggtc tggtctcccc atgcgggcca ccagaggccc tgcagccttc agtcgcttga
     3541 aggggtaatg gcgcttccac tcacaaacat ggcggacaga gcgtgtgaac gagatgaaca
     3601 gcccctcaaa aatatggccg ccgaggctgg acggccgtgc cccagcagca ccgcctccgc
     3661 gccccacgtg atctctcgcc gggcacagcg ctgaccgcgg aggtccaacc ggaagaatgt
     3721 ccggattgga cattcggaag agggcccgcc ttccctgggg aatctctgcg cacgcgcaga
     3781 acgcttcgac caatgaaaac acaggaagcc gtccgcgcaa ccgcgttgcg tcacttctgc
     3841 cgcccctgtt tcaaggtata tagccgtaga cggaacttcg cctttctctc ggccttagcg
     3901 ccattttttt gggtgagtgt tttttggttc ctgcgttggg attccgtgta caatccatag
     3961 acatctgacc tcggcactta gcatcatcac agcaaactaa ctgtagcctt tctctctttc
     4021 cctgtagaaa cctctgcacc tgaggccacc aaactagcca ccatggccaa gttgaccagt
     4081 gccgttccgg tgctcaccgc gcgcgacgtc gccggagcgg tcgagttctg gaccgaccgg
     4141 ctcgggttct cccgggactt cgtggaggac gacttcgccg gtgtggtccg ggacgacgtg
     4201 accctgttca tcagcgcggt ccaggaccag gtggtgccgg acaacaccct ggcctgggtg
     4261 tgggtgcgcg gcctggacga gctgtacgcc gagtggtcgg aggtcgtgtc cacgaacttc
     4321 cgggacgcct ccgggccggc catgaccgag atcggcgagc agccgtgggg gcgggagttc
     4381 gccctgcgcg acccggccgg caactgcgtg cacttcgtgg ccgaggagca ggactgattc
     4441 gaaggcctgt cgtgaagctt ggggatcaat tctctagagc tcgctgatca gcctcgactg
     4501 tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg
     4561 aaggtgcccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag
     4621 gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga
     4681 caatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag
     4741 ctgggcctac tagctactcg ggacccctta ccgaaacatc gccgcattct gcagaggagt
     4801 cgagtgtatg taaacttctg acccactggg aatgtgatga aagaaataaa agctgaaatg
     4861 aatcattctc tctactatta ttctgatatt tcacattctt aaaataaagt ggtgatccta
     4921 actgacctaa gacagggaat ttttactagg attaaatgtc aggaattgtg aaaaagtgag
     4981 tttaaatgta tttggctaag gtgtatgtaa acttccgact tcaactgtat agggatccgc
     5041 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     5101 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     5161 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     5221 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     5281 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     5341 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     5401 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     5461 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     5521 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     5581 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     5641 cggctacact agaagaacag tatttggtat ctgcgctct
//
