LOCUS       sequence-422617-        3310 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ATc-inducible himar1C9 transposase expression - cam resistance.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3310)
  AUTHORS   English MA, Alcantar MA, Collins JJ
  TITLE     A self-propagating, barcoded transposon system for the dynamic 
            rewiring of genomic networks.
  JOURNAL   Mol Syst Biol. 2023 Jun 12;19(6):e11398. doi: 
            10.15252/msb.202211398. Epub 2023 Mar 27.
  PUBMED    36970845
REFERENCE   2  (bases 1 to 3310)
  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: "Mol Syst 
            Biol."; date: "2023-06-12"; volume: "19(6):e11398. doi"; pages: " 
            10.15252/msb.202211398. Epub 2023 Mar 27"
COMMENT     Sequence Label: pHelper_TetR/pTet-Tnpase (camR)
FEATURES             Location/Qualifiers
     source          1..3310
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(1..546)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     complement(81..100)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             662..1321
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(728..747)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      1392..1426
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     CDS             complement(1431..2057)
                     /codon_start=1
                     /gene="tetR"
                     /product="tetracycline repressor TetR"
                     /label=TetR
                     /note="TetR binds to the tetracycline operator tetO to 
                     inhibit transcription. This inhibition can be relieved by 
                     adding tetracycline or doxycycline."
                     /translation="MMSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWH
                     VKNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGT
                     RPTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPT
                     TDSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     primer_bind     1962..1981
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     promoter        2073..2128
                     /gene="tetR"
                     /label=tetR/tetA promoters
                     /note="overlapping promoters for bacterial tetR and tetA"
     protein_bind    2109..2127
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     RBS             2146..2157
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
ORIGIN
        1 tttccatagg ctccgccccc ctgacaagca tcacgaaatc tgacgctcaa atcagtggtg
       61 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggcggct ccctcgtgcg
      121 ctctcctgtt cctgcctttc ggtttaccgg tgtcattccg ctgttatggc cgcgtttgtc
      181 tcattccacg cctgacactc agttccgggt aggcagttcg ctccaagctg gactgtatgc
      241 acgaaccccc cgttcagtcc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
      301 acccggaaag acatgcaaaa gcaccactgg cagcagccac tggtaattga tttagaggag
      361 ttagtcttga agtcatgcgc cggttaaggc taaactgaaa ggacaagttt tggtgactgc
      421 gctcctccaa gccagttacc tcggttcaaa gagttggtag ctcagagaac cttcgaaaaa
      481 ccgccctgca aggcggtttt ttcgttttca gagcaagaga ttacgcgcag accaaaacga
      541 tctcaagaag atcatcttga tcgggcacgt aagaggttcc aactttcacc ataatgaaat
      601 aagatcacta ccgggcgtat tttttgagtt atcgagattt tcaggagcta aggaagctaa
      661 aatggagaaa aaaatcactg gatataccac cgttgatata tcccaatggc atcgtaaaga
      721 acattttgag gcatttcagt cagttgctca atgtacctat aaccagaccg ttcagctgga
      781 tattacggcc tttttaaaga ccgtaaagaa aaataagcac aagttttatc cggcctttat
      841 tcacattctt gcccgcctga tgaatgctca tccggaattt cgtatggcaa tgaaagacgg
      901 tgagctggtg atatgggata gtgttcaccc ttgttacacc gttttccatg agcaaactga
      961 aacgttttca tcgctctgga gtgaatacca cgacgatttc cggcagtttc tacacatata
     1021 ttcgcaagat gtggcgtgtt acggtgaaaa cctggcctat ttccctaaag ggtttattga
     1081 gaatatgttt ttcgtttcag ccaatccctg ggtgagtttc accagttttg atttaaacgt
     1141 ggccaatatg gacaacttct tcgcccccgt tttcaccatg ggcaaatatt atacgcaagg
     1201 cgacaaggtg ctgatgccgc tggcgattca ggttcatcat gccgtttgtg atggcttcca
     1261 tgtcggcaga atgcttaatg aattacaaca gtactgcgat gagtggcagg gcggggcgta
     1321 atttgatatc gagctcgctt ggactcctgt tgatagatcc agtaatgacc tcagaactcc
     1381 atctggattt gttcagaacg ctcggttgcc gccgggcgtt ttttattggt ttaagaccca
     1441 ctttcacatt taagttgttt ttctaatccg catatgatca attcaaggcc gaataagaag
     1501 gctggctctg caccttggtg atcaaataat tcgatagctt gtcgtaataa tggcggcata
     1561 ctatcagtag taggtgtttc cctttcttct ttagcgactt gatgctcttg atcttccaat
     1621 acgcaaccta aagtaaaatg ccccacagcg ctgagtgcat ataatgcatt ctctagtgaa
     1681 aaaccttgtt ggcataaaaa ggctaattga ttttcgagag tttcatactg tttttctgta
     1741 ggccgtgtac ctaaatgtac ttttgctcca tcgcgatgac ttagtaaagc acatctaaaa
     1801 cttttagcgt tattacgtaa aaaatcttgc cagctttccc cttctaaagg gcaaaagtga
     1861 gtatggtgcc tatctaacat ctcaatggct aaggcgtcga gcaaagcccg cttatttttt
     1921 acatgccaat acaatgtagg ctgctctaca cctagcttct gggcgagttt acgggttgtt
     1981 aaaccttcga ttccgacctc attaagcagc tctaatgcgc tgttaatcac tttactttta
     2041 tctaatctag acatcattaa ttcctaattt ttgttgacac tctatcgttg atagagttat
     2101 tttaccactc cctatcagtg atagagaaaa gaattcaaaa gatctaaaga ggagaaagga
     2161 tctatggaaa aaaaggaatt tcgtgttttg ataaaatact gttttctgaa gggaaaaaat
     2221 acagtggaag caaaaacttg gcttgataat gagtttccgg actctgcccc ggggaaatca
     2281 acaataattg attggtatgc aaaattcaag cgtggtgaaa tgagcacgga ggacggtgaa
     2341 cgcagtggac gcccgaaaga ggtggttacc gacgaaaaca tcaaaaaaat ccacaaaatg
     2401 attttgaatg accgtaaaat gaagttgatc gagatagcag aggccttaaa gatatcaaag
     2461 gaacgtgttg gtcatatcat tcatcaatat ttggatatgc ggaagctctg tgcgaaatgg
     2521 gtgccgcgcg agctcacatt tgaccaaaaa caacgacgtg ttgatgattc taagcggtgt
     2581 ttgcagctgt taactcgtaa tacacccgag tttttccgtc gatatgtgac aatggatgaa
     2641 acatggctcc atcactacac tcctgagtcc aatcgacagt cggctgagtg gacagcgacc
     2701 ggtgaaccgt ctccgaagcg tggaaagact caaaagtccg ctggcaaagt aatggcctct
     2761 gttttttggg atgcgcatgg aataattttt atcgattatc ttgagaaggg aaaaaccatc
     2821 aacagtgact attatatggc gttattggag cgtttgaagg tcgaaatcgc ggcaaaacgg
     2881 ccccacatga agaagaaaaa agtgttgttc caccaagaca acgcaccgtg ccacaagtca
     2941 ttgagaacga tggcaaaaat tcatgaattg ggcttcgaat tgcttcccca cccgccgtat
     3001 tctccagatc tggcccccag cgactttttc ttgttctcag acctcaaaag gatgctcgca
     3061 gggaaaaaat ttggctgcaa tgaagaggtg atcgccgaaa ctgaggccta ttttgaggca
     3121 aaaccgaagg agtactacca aaatggtatc aaaaaattgg aaggtcgtta taatcgttgt
     3181 atcgctcttg aagggaacta tgttgaataa ctgtaacaga gcattagcgc aaggtgattt
     3241 ttgtcttctt gcgctaattt tttcaggaag atacttaata gggaagtgag agggccgcgg
     3301 caaagccgtt
//
