LOCUS       sequence-422618-        3787 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ATc-inducible himar1C9 transposase expression - carb/amp resistance.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3787)
  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 3787)
  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 (carbR)
FEATURES             Location/Qualifiers
     source          1..3787
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(7..633)
                     /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     538..557
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     promoter        649..704
                     /gene="tetR"
                     /label=tetR/tetA promoters
                     /note="overlapping promoters for bacterial tetR and tetA"
     protein_bind    685..703
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     RBS             722..733
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     terminator      1810..1856
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(1893..1910)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2027..2572)
                     /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(2107..2126)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(2686..2780)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(join(2801..3592,3593..3661))
                     /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     3424..3443
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3662..3764)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
ORIGIN
        1 gacgtcttaa gacccacttt cacatttaag ttgtttttct aatccgcata tgatcaattc
       61 aaggccgaat aagaaggctg gctctgcacc ttggtgatca aataattcga tagcttgtcg
      121 taataatggc ggcatactat cagtagtagg tgtttccctt tcttctttag cgacttgatg
      181 ctcttgatct tccaatacgc aacctaaagt aaaatgcccc acagcgctga gtgcatataa
      241 tgcattctct agtgaaaaac cttgttggca taaaaaggct aattgatttt cgagagtttc
      301 atactgtttt tctgtaggcc gtgtacctaa atgtactttt gctccatcgc gatgacttag
      361 taaagcacat ctaaaacttt tagcgttatt acgtaaaaaa tcttgccagc tttccccttc
      421 taaagggcaa aagtgagtat ggtgcctatc taacatctca atggctaagg cgtcgagcaa
      481 agcccgctta ttttttacat gccaatacaa tgtaggctgc tctacaccta gcttctgggc
      541 gagtttacgg gttgttaaac cttcgattcc gacctcatta agcagctcta atgcgctgtt
      601 aatcacttta cttttatcta atctagacat cattaattcc taatttttgt tgacactcta
      661 tcgttgatag agttatttta ccactcccta tcagtgatag agaaaagaat tcaaaagatc
      721 taaagaggag aaaggatcta tggaaaaaaa ggaatttcgt gttttgataa aatactgttt
      781 tctgaaggga aaaaatacag tggaagcaaa aacttggctt gataatgagt ttccggactc
      841 tgccccgggg aaatcaacaa taattgattg gtatgcaaaa ttcaagcgtg gtgaaatgag
      901 cacggaggac ggtgaacgca gtggacgccc gaaagaggtg gttaccgacg aaaacatcaa
      961 aaaaatccac aaaatgattt tgaatgaccg taaaatgaag ttgatcgaga tagcagaggc
     1021 cttaaagata tcaaaggaac gtgttggtca tatcattcat caatatttgg atatgcggaa
     1081 gctctgtgcg aaatgggtgc cgcgcgagct cacatttgac caaaaacaac gacgtgttga
     1141 tgattctaag cggtgtttgc agctgttaac tcgtaataca cccgagtttt tccgtcgata
     1201 tgtgacaatg gatgaaacat ggctccatca ctacactcct gagtccaatc gacagtcggc
     1261 tgagtggaca gcgaccggtg aaccgtctcc gaagcgtgga aagactcaaa agtccgctgg
     1321 caaagtaatg gcctctgttt tttgggatgc gcatggaata atttttatcg attatcttga
     1381 gaagggaaaa accatcaaca gtgactatta tatggcgtta ttggagcgtt tgaaggtcga
     1441 aatcgcggca aaacggcccc acatgaagaa gaaaaaagtg ttgttccacc aagacaacgc
     1501 accgtgccac aagtcattga gaacgatggc aaaaattcat gaattgggct tcgaattgct
     1561 tccccacccg ccgtattctc cagatctggc ccccagcgac tttttcttgt tctcagacct
     1621 caaaaggatg ctcgcaggga aaaaatttgg ctgcaatgaa gaggtgatcg ccgaaactga
     1681 ggcctatttt gaggcaaaac cgaaggagta ctaccaaaat ggtatcaaaa aattggaagg
     1741 tcgttataat cgttgtatcg ctcttgaagg gaactatgtt gaataactcg agtaaggatc
     1801 tccaggcatc aaataaaacg aaaggctcag tcgaaagact gggcctttcg ttttatccga
     1861 atacgggttc ttgcatggat atattccgct tcctcgctca ctgactcgct acgctcggtc
     1921 gttcgactgc ggcgagcgga aatggcttac gaacggggcg gagatttcct ggaagatgcc
     1981 aggaagatac ttaataggga agtgagaggg ccgcggcaaa gccgtttttc cataggctcc
     2041 gcccccctga caagcatcac gaaatctgac gctcaaatca gtggtggcga aacccgacag
     2101 gactataaag ataccaggcg tttccccctg gcggctccct cgtgcgctct cctgttcctg
     2161 cctttcggtt taccggtgtc attccgctgt tatggccgcg tttgtctcat tccacgcctg
     2221 acactcagtt ccgggtaggc agttcgctcc aagctggact gtatgcacga accccccgtt
     2281 cagtccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggaaagacat
     2341 gcaaaagcac cactggcagc agccactggt aattgattta gaggagttag tcttgaagtc
     2401 atgcgccggt taaggctaaa ctgaaaggac aagttttggt gactgcgctc ctccaagcca
     2461 gttacctcgg ttcaaagagt tggtagctca gagaaccttc gaaaaaccgc cctgcaaggc
     2521 ggttttttcg ttttcagagc aagagattac gcgcagacca aaacgatctc aagaagatca
     2581 tcttattaat cagataaaat atttctagat ttcagtgcaa tttatctctt caaatgtagc
     2641 acctgaagtc agccccatac gatataagtt gttactagtg cttggattct caccaataaa
     2701 aaacgcccgg cggcaaccga gcgttctgaa caaatccaga tggagttctg aggtcattac
     2761 tggatctatc aacaggagtc caagcgagct cgatatcaaa ttaccaatgc ttaatcagtg
     2821 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     2881 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     2941 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     3001 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     3061 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     3121 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     3181 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagttcc ttcggtcctc
     3241 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     3301 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     3361 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     3421 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     3481 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     3541 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     3601 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     3661 ttttagcttc cttagctcct gaaaatctcg ataactcaaa aaatacgccc ggtagtgatc
     3721 ttatttcatt atggtgaaag ttggaacctc ttacgtgccg atcaacgtct cattttcgcc
     3781 agatatc
//
