LOCUS       sequence-352286-        3558 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3558)
  TITLE     Transposon cloning vectors
REFERENCE   2  (bases 1 to 3558)
  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: pT2/HB
FEATURES             Location/Qualifiers
     source          1..3558
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(147..735)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(227..246)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(906..1697,1698..1766))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAAKLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     1529..1548
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1767..1871)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      1898..2353
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1985..2004)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2195..2216
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2479..2501
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2493..2510
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2494..2510
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2520..2539
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2520..2538
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3264..3284)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3264..3282)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3303..3319)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3303..3319)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3316..3338)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3327..3343
                     /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(3351..3357,3358..3375,3376..3381))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3396..3417
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3534..3551)
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
       61 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
      121 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
      181 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
      241 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
      301 ctgtccgcct ttctcccttc gggaaccgtg gcgctttctc atagctcacg ctgtaggtat
      361 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
      421 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
      481 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
      541 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
      601 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
      661 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
      721 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
      781 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
      841 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
      901 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
      961 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     1021 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     1081 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     1141 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     1201 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     1261 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     1321 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaacttggc cgcagtgtta
     1381 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     1441 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     1501 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     1561 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     1621 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     1681 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     1741 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     1801 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     1861 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg cgccctgtag cggcgcatta
     1921 agcgcggcgg gagtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
     1981 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     2041 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     2101 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     2161 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     2221 acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc
     2281 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta
     2341 acgcttacaa tttccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc
     2401 gggcctcttc gctattacgc cagctggcga aagggggatg tgctgcaagg cgattaagtt
     2461 gggtaacgcc agggttttcc cagtcacgac gttgtaaaac gacggccagt gagcgcgcgt
     2521 aatacgactc actatagggc gaattggagc tcggatccct atacagttga agtcggaagt
     2581 ttacatacac ttaagttgga gtcattaaaa ctcgtttttc aactactcca caaatttctt
     2641 gttaacaaac aatagttttg gcaagtcagt taggacatct actttgtgca tgacacaagt
     2701 catttttcca acaattgttt acagacagat tatttcactt ataattcact gtatcacaat
     2761 tccagtgggt cagaagttta catacactaa gttgactgtg cctttaaaca gcttggaaaa
     2821 ttccagaaaa tgatgtcatg gctttagaag cttgatatcc atggaattca ctagtgcgcg
     2881 cggccgctct agatggccag atctagcttg tggaaggcta ctcgaaatgt ttgacccaag
     2941 ttaaacaatt taaaggcaat gctaccaaat actaattgag tgtatgtaaa cttctgaccc
     3001 actgggaatg tgatgaaaga aataaaagct gaaatgaatc attctctcta ctattattct
     3061 gatatttcac attcttaaaa taaagtggtg atcctaactg acctaagaca gggaattttt
     3121 actaggatta aatgtcagga attgtgaaaa agtgagttta aatgtatttg gctaaggtgt
     3181 atgtaaactt ccgacttcaa ctgtataggg atcctctagc tagagtcgac ctcgaggggg
     3241 ggcccggtac ccagcttttg ttccctttag tgagggttaa ttgcgcgctt ggcgtaatca
     3301 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
     3361 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
     3421 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
     3481 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     3541 actgactcgc tgcgctcg
//
