LOCUS       sequence-309397-        5227 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gal4-inducible tag blue fluorescent protein (tBFP) in pSB-Bleo Gal4 
            Sleeping Beauty backbone..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5227)
  TITLE     Methylxanthine Synthesis in Human Cells
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5227)
  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            "Unpublished"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 tBFP
FEATURES             Location/Qualifiers
     source          1..5227
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     371..395
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             489..1190
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /label=TagBFP
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALK
                     LVGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYC
                     DLPSKLGHKLN"
     polyA_signal    1239..1360
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1250..1269)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1304..1323
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     regulatory      2114..2123
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2120..2494
                     /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(2549..2566)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     primer_bind     complement(3101..3118)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3272..3860)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3352..3371)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4031..4822,4823..4891))
                     /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     4654..4673
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4892..4996)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 catctacttt gtgcatgaca caagtcattt ttccaacaat tgtttacaga cagattattt
       61 cacttataat tcactgtatc acaattccag tgggtcagaa gtttacatac actaagttcg
      121 actcctctgc agaatgcggc gatgtttcgg taaggggtcc gctatctagt gagctcttac
      181 gcgtgctagc cttgatatcg aattcggagc actgtcctcc gaacgtcgga gcactgtcct
      241 ccgaacgtcg gagcactgtc ctccgaacgt cggagcactg tcctccgaac ggagcatgtc
      301 ctccgaacgt cggagcactg tcctccgaac gactagttag gcgtgtacgg tgggaggcct
      361 atataagcag agctcgttta gtgaaccgtc agatcgcctg gagacgccat ccacgctgtt
      421 ttgacctcca tagaagacac cgggaccgat ccagcctctc gacattcgtt ggatcggcct
      481 ctgaggccat gagcgagctg attaaggaga acatgcacat gaagctgtac atggagggca
      541 ccgtggacaa ccatcacttc aagtgcacat ccgagggcga aggcaagccc tacgagggca
      601 cccagaccat gagaatcaag gtggtcgagg gcggccctct ccccttcgcc ttcgacatcc
      661 tggctactag cttcctctac ggcagcaaga ccttcatcaa ccacacccag ggcatccccg
      721 acttcttcaa gcagtccttc cctgagggct tcacatggga gagagtcacc acatacgaag
      781 acgggggcgt gctgaccgct acccaggaca ccagcctcca ggacggctgc ctcatctaca
      841 acgtcaagat cagaggggtg aacttcacat ccaacggccc tgtgatgcag aagaaaacac
      901 tcggctggga ggccttcacc gagacgctgt accccgctga cggcggcctg gaaggcagaa
      961 acgacatggc cctgaagctc gtgggcggga gccatctgat cgcaaacatc aagaccacat
     1021 atagatccaa gaaacccgct aagaacctca agatgcctgg cgtctactat gtggactaca
     1081 gactggaaag aatcaaggag gccaacaacg agacctacgt cgagcagcac gaggtggcag
     1141 tggccagata ctgcgacctc cctagcaaac tggggcacaa gcttaattaa gatccttgac
     1201 tattcgaagg cctgtcaggc cgcttcgagc agacatgata agatacattg atgagtttgg
     1261 acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat
     1321 tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca
     1381 ttttatgttt caggttcagg gggaggtgtg ggaggttttt taaagcaagt aaaacctcta
     1441 caaatgtggt aaaatcgata aggatccgtc gaccggtacc taggcgagac cctgtcacaa
     1501 aataaagtaa gcccggactg agtgcggaaa ggcgggcctg gcgggtctgg tctccccatg
     1561 cgggccacca gaggccctgc agccttcagt cgcttgaagg ggtaatggcg cttccactca
     1621 caaacatggc ggacagagcg tgtgaacgag atgaacagcc cctcaaaaat atggccgccg
     1681 aggctggacg gccgtgcccc agcagcaccg cctccgcgcc ccacgtgatc tctcgccggg
     1741 cacagcgctg accgcggagg tccaaccgga agaatgtccg gattggacat tcggaagagg
     1801 gcccgccttc cctggggaat ctctgcgcac gcgcagaacg cttcgaccaa tgaaaacaca
     1861 ggaagccgtc cgcgcaaccg cgttgcgtca cttctgccgc ccctgtttca aggtatatag
     1921 ccgtagacgg aacttcgcct ttctctcggc cttagcgcca tttttttggg tgagtgtttt
     1981 ttggttcctg cgttgggatt ccgtgtacaa tccatagaca tctgacctcg gcacttagca
     2041 tcatcacagc aaactaactg tagcctttct ctctttccct gtagaaacct ctgcacctga
     2101 ggccaccaaa ctagccacca tggccaagtt gaccagtgcc gttccggtgc tcaccgcgcg
     2161 cgacgtcgcc ggagcggtcg agttctggac cgaccggctc gggttctccc gggacttcgt
     2221 ggaggacgac ttcgccggtg tggtccggga cgacgtgacc ctgttcatca gcgcggtcca
     2281 ggaccaggtg gtgccggaca acaccctggc ctgggtgtgg gtgcgcggcc tggacgagct
     2341 gtacgccgag tggtcggagg tcgtgtccac gaacttccgg gacgcctccg ggccggccat
     2401 gaccgagatc ggcgagcagc cgtgggggcg ggagttcgcc ctgcgcgacc cggccggcaa
     2461 ctgcgtgcac ttcgtggccg aggagcagga ctgattcgaa ggcctgtcgt gaagcttggg
     2521 gatcaattct ctagagctcg ctgatcagcc tcgactgtgc cttctagttg ccagccatct
     2581 gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccccact gtcctttcct
     2641 aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg
     2701 gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg
     2761 cggtgggctc tatggcttct gaggcggaaa gaaccagctg ggcctactag ctactcggga
     2821 ccccttaccg aaacatcgcc gcattctgca gaggagtcga gtgtatgtaa acttctgacc
     2881 cactgggaat gtgatgaaag aaataaaagc tgaaatgaat cattctctct actattattc
     2941 tgatatttca cattcttaaa ataaagtggt gatcctaact gacctaagac agggaatttt
     3001 tactaggatt aaatgtcagg aattgtgaaa aagtgagttt aaatgtattt ggctaaggtg
     3061 tatgtaaact tccgacttca actgtatagg gatccgcttc ctcgctcact gactcgctgc
     3121 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     3181 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     3241 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     3301 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     3361 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     3421 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     3481 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     3541 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     3601 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     3661 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     3721 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     3781 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     3841 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     3901 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     3961 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     4021 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     4081 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     4141 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     4201 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     4261 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     4321 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     4381 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     4441 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     4501 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     4561 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     4621 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     4681 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     4741 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     4801 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     4861 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     4921 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     4981 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgatgcggtg tgaaataccg
     5041 cacagatgcg taaggagaaa ataccgcatc aggaaattgt aagcgttaat attttgttaa
     5101 aattggatcc ctatacagtt gaagtcggaa gtttacatac acttaagttg gagtcattaa
     5161 aactcgtttt tcaactactc cacaaatttc ttgttaacaa acaatagttt tggcaagtca
     5221 gttagga
//
