LOCUS       sequence-309370-        4638 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Empty backbone encoding constitutive bleomycin resistance with an 
            inducible STAT3 promoter to allow for an insert in a Sleeping Beauty
            backbone..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4638)
  TITLE     Methylxanthine Synthesis in Human Cells
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4638)
  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            "Unpublished"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 STAT3
FEATURES             Location/Qualifiers
     source          1..4638
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(28..45)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(199..787)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(279..298)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(958..1749,1750..1818))
                     /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     1581..1600
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1819..1923)
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2426..2464
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2461..2485
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      2575..2584
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    2804..2925
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2815..2834)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2869..2888
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     regulatory      3679..3688
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3685..4059
                     /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(4114..4131)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
ORIGIN
        1 gacttcaact gtatagggat ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
       61 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
      121 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
      181 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
      241 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
      301 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
      361 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
      421 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
      481 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
      541 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
      601 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
      661 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
      721 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
      781 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
      841 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
      901 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
      961 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     1021 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     1081 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     1141 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     1201 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     1261 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     1321 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     1381 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     1441 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     1501 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     1561 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     1621 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     1681 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     1741 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     1801 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     1861 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     1921 gcgcacattt ccccgaaaag tgccacctga tgcggtgtga aataccgcac agatgcgtaa
     1981 ggagaaaata ccgcatcagg aaattgtaag cgttaatatt ttgttaaaat tggatcccta
     2041 tacagttgaa gtcggaagtt tacatacact taagttggag tcattaaaac tcgtttttca
     2101 actactccac aaatttcttg ttaacaaaca atagttttgg caagtcagtt aggacatcta
     2161 ctttgtgcat gacacaagtc atttttccaa caattgttta cagacagatt atttcactta
     2221 taattcactg tatcacaatt ccagtgggtc agaagtttac atacactaag ttcgactcct
     2281 ctgcagaatg cggcgatgtt tcggtaaggg gtccgctatc tagtgagctc ttacgcgtgc
     2341 tagccgtcga catttcccgt aaatcgtcga gtcgacattt cccgtaaatc gtcgacctgc
     2401 aggtcgagct cggtacccgg gtcgaggtag gcgtgtacgg tgggaggcct atataagcag
     2461 agctcgttta gtgaaccgtc agatcgcctg gagacgccat ccacgctgtt ttgacctcca
     2521 tagaagacac cgggaccgat ccagcctccg cgggaattca agctggcctc tgaggccacc
     2581 atggctcgag tttcgaactc aattatactg aattgtttca tcatagggtt caacattgtg
     2641 ataaatcagg acttcaatgt ctagtgttca gtaaatacct tgtagggctc tccaaaaagg
     2701 ggaatctttt caggaaatgc ctctttctct tggtgggctt cctggttgcg tctttccttc
     2761 tcactaattc gaaggcctgt caggccgctt cgagcagaca tgataagata cattgatgag
     2821 tttggacaaa ccacaactag aatgcagtga aaaaaatgct ttatttgtga aatttgtgat
     2881 gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa caacaattgc
     2941 attcatttta tgtttcaggt tcagggggag gtgtgggagg ttttttaaag caagtaaaac
     3001 ctctacaaat gtggtaaaat cgataaggat ccgtcgaccg gtacctaggc gagaccctgt
     3061 cacaaaataa agtaagcccg gactgagtgc ggaaaggcgg gcctggcggg tctggtctcc
     3121 ccatgcgggc caccagaggc cctgcagcct tcagtcgctt gaaggggtaa tggcgcttcc
     3181 actcacaaac atggcggaca gagcgtgtga acgagatgaa cagcccctca aaaatatggc
     3241 cgccgaggct ggacggccgt gccccagcag caccgcctcc gcgccccacg tgatctctcg
     3301 ccgggcacag cgctgaccgc ggaggtccaa ccggaagaat gtccggattg gacattcgga
     3361 agagggcccg ccttccctgg ggaatctctg cgcacgcgca gaacgcttcg accaatgaaa
     3421 acacaggaag ccgtccgcgc aaccgcgttg cgtcacttct gccgcccctg tttcaaggta
     3481 tatagccgta gacggaactt cgcctttctc tcggccttag cgccattttt ttgggtgagt
     3541 gttttttggt tcctgcgttg ggattccgtg tacaatccat agacatctga cctcggcact
     3601 tagcatcatc acagcaaact aactgtagcc tttctctctt tccctgtaga aacctctgca
     3661 cctgaggcca ccaaactagc caccatggcc aagttgacca gtgccgttcc ggtgctcacc
     3721 gcgcgcgacg tcgccggagc ggtcgagttc tggaccgacc ggctcgggtt ctcccgggac
     3781 ttcgtggagg acgacttcgc cggtgtggtc cgggacgacg tgaccctgtt catcagcgcg
     3841 gtccaggacc aggtggtgcc ggacaacacc ctggcctggg tgtgggtgcg cggcctggac
     3901 gagctgtacg ccgagtggtc ggaggtcgtg tccacgaact tccgggacgc ctccgggccg
     3961 gccatgaccg agatcggcga gcagccgtgg gggcgggagt tcgccctgcg cgacccggcc
     4021 ggcaactgcg tgcacttcgt ggccgaggag caggactgat tcgaaggcct gtcgtgaagc
     4081 ttggggatca attctctaga gctcgctgat cagcctcgac tgtgccttct agttgccagc
     4141 catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc ccactgtcct
     4201 ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg
     4261 gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg
     4321 ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc agctgggcct actagctact
     4381 cgggacccct taccgaaaca tcgccgcatt ctgcagagga gtcgagtgta tgtaaacttc
     4441 tgacccactg ggaatgtgat gaaagaaata aaagctgaaa tgaatcattc tctctactat
     4501 tattctgata tttcacattc ttaaaataaa gtggtgatcc taactgacct aagacaggga
     4561 atttttacta ggattaaatg tcaggaattg tgaaaaagtg agtttaaatg tatttggcta
     4621 aggtgtatgt aaacttcc
//
