LOCUS       sequence-12574-e        3537 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  natural linear DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 3537)
  TITLE     Transposon cloning vectors
REFERENCE   2  (bases 1 to 3537)
  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"
COMMENT     Sequence Label: pT/HB
FEATURES             Location/Qualifiers
     source          1..3537
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     rep_origin      3..458
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(90..109)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     300..321
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     584..606
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     598..615
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     599..615
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     622..641
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        622..640
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(1348..1366)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1387..1403)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1387..1403)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1400..1422)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1411..1427
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(1435..1441,1442..1459,1460..1465))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1480..1501
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1618..1635)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1789..2377)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1869..1888)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2548..3339,3340..3408))
                     /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     3171..3190
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3409..3513)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tgacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
       61 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
      121 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
      181 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg
      241 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
      301 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
      361 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
      421 taacgcgaat tttaacaaaa tattaacgct tacaatttcc attcgccatt caggctgcgc
      481 aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg
      541 ggatgtgctg caaggcgatt aagttgggta acgccagggt tttcccagtc acgacgttgt
      601 aaaacgacgg ccagtgaatt gtaatacgac tcactatagg gcgaattgga gctcggtacc
      661 ctacagttga agtcggaagt ttacatacac ttaagttgga gtcattaaaa ctcgtttttc
      721 aactacacca caaatttctt gttaacaaac aatagttttg gcaagtcagt taggacatct
      781 actttgtgca tgacacaagt catttttcca acaattgttt acagacagat tatttcactt
      841 ataattcact gtatcacaat tccagtgggt cagaagttta catacactaa gttgactgtg
      901 cctttaaaca gcttggaaaa ttccagaaaa tgatgtcatg gctttagaag cttgatatcg
      961 aattcactag tgcggccgct ctagatctag cttgtggaag gctactcgaa atgtttgacc
     1021 caagttaaac aatttaaagg caatgctacc aaatactaat tgagtgtatg ttaacttctg
     1081 acccactggg aatgtgatga aagaaataaa agctgaaatg aatcattctc tctactatta
     1141 ttctgatatt tcacattctt aaaataaagt ggtgatccta actgacctta agacagggaa
     1201 tctttactcg gattaaatgt caggaattgt gaaaaagtga gtttaaatgt atttggctaa
     1261 ggtgtatgta aacttccgac ttcaactgta ggggatcctc tagctagagt cgacctcgag
     1321 ggggggcccg gtacccagct tttgttccct ttagtgaggg ttaatttcga gcttggcgta
     1381 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
     1441 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
     1501 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     1561 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     1621 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     1681 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     1741 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     1801 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     1861 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     1921 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     1981 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     2041 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     2101 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     2161 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     2221 cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     2281 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     2341 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     2401 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     2461 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     2521 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     2581 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     2641 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     2701 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     2761 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     2821 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     2881 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     2941 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     3001 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     3061 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     3121 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     3181 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     3241 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     3301 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     3361 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     3421 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     3481 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacc
//
