LOCUS       sequence-401409-        6525 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Arabinose inducible expression vector expressing retron Rcat-Sen2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6525)
  AUTHORS   Bobonis J, Yang ALJ, Voogdt CGP, Typas A
  TITLE     TAC-TIC, a high-throughput genetics method to identify triggers or 
            blockers of bacterial toxin-antitoxin systems.
  JOURNAL   Nat Protoc. 2024 May 9. doi: 10.1038/s41596-024-00988-y.
  PUBMED    38724726
REFERENCE   2  (bases 1 to 6525)
  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: "Nat Protoc.
            2024 May 9. doi: 10.1038/s41596-024-00988-y."
COMMENT     Sequence Label: pJB37
FEATURES             Location/Qualifiers
     source          1..6525
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(254..1045)
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     CDS             complement(2375..3325)
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
     rep_origin      3373..3595
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     CDS             complement(4331..5209)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        5236..5520
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     5443..5462
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
ORIGIN
        1 aagcttagta aagccctcgc tagattttaa tgcggatgtt gcgattactt cgccaactat
       61 tgcgataaca agaaaaagcc agcctttcat gatatatctc ccaatttgtg tagggcttat
      121 tatgcacgct taaaaataat aaaagcagac ttgacctgat agtttggctg tgagcaatta
      181 tgtgcttagt gcatctaacg cttgagttaa gccgcgccgc gaagcggcgt cggcttgaac
      241 gaattgttag acattatttg ccgactacct tggtgatctc gcctttcacg tagtggacaa
      301 attcttccaa ctgatctgcg cgcgaggcca agcgatcttc ttcttgtcca agataagcct
      361 gtctagcttc aagtatgacg ggctgatact gggccggcag gcgctccatt gcccagtcgg
      421 cagcgacatc cttcggcgcg attttgccgg ttactgcgct gtaccaaatg cgggacaacg
      481 taagcactac atttcgctca tcgccagccc agtcgggcgg cgagttccat agcgttaagg
      541 tttcatttag cgcctcaaat agatcctgtt caggaaccgg atcaaagagt tcctccgccg
      601 ctggacctac caaggcaacg ctatgttctc ttgcttttgt cagcaagata gccagatcaa
      661 tgtcgatcgt ggctggctcg aagatacctg caagaatgtc attgcgctgc cattctccaa
      721 attgcagttc gcgcttagct ggataacgcc acggaatgat gtcgtcgtgc acaacaatgg
      781 tgacttctac agcgcggaga atctcgctct ctccagggga agccgaagtt tccaaaaggt
      841 cgttgatcaa agctcgccgc gttgtttcat caagccttac ggtcaccgta accagcaaat
      901 caatatcact gtgtggcttc aggccgccat ccactgcgga gccgtacaaa tgtacggcca
      961 gcaacgtcgg ttcgagatgg cgctcgatga cgccaactac ctctgatagt tgagtcgata
     1021 cttcggcgat caccgcttcc ctcatgatgt ttaactttgt tttagggcga ctgccctgct
     1081 gcgtaacatc gttgctgctc cataacatca aacatcgacc cacggcgtaa cgcgcttgct
     1141 gcttggatgc ccgaggcata gactgtaccc caaaaaaaca gtcataacaa gccatgaaaa
     1201 ccgccactgc gccgttacca ccgctgcgtt cggtcaaggt tctggaccag ttgcgtgagc
     1261 gcatacgcta cttgcattac agcttacgaa ccgaacaggc ttatgtccac tgggttcgtg
     1321 ccttcatccg tttccacggt gtgcgtcacc cggcaacctt gggcagcagc gaagtcgagg
     1381 catttctgtc ctggctggcg aacgagcgca aggtttcggt ctccacgcat cgtcaggcat
     1441 tggcggcctt gctgttcttc tacggcaagg tgctgtgcac ggatctgccc tggcttcagg
     1501 agatcggaag acctcggccg ccacctctgg tgactttatc cgtaaataat ttaacccact
     1561 ccacaaaaag gctcaacagg ttggtggttc tcaccaccaa aagcaccaca ccccacgcaa
     1621 aaacaagttt ttgctgattt ttctttataa atagagtgtt atgaaaaatt agtttctctt
     1681 actctcttta tgatatttaa aaaagcggtg tcggcgcggc tacaacaacg cgccgacacc
     1741 gttttgtagg ggtggtactg actattttta taaaaaacat tattttatat taggggtgct
     1801 gctagcggcg cggtgtgttt ttttatagga taccgctagg ggcgctgcat taattcccac
     1861 gggttttgct gcccgcaaac gggctgttct ggtgttgcta gtttgttatc agaatcgcag
     1921 atccggcttc agccggtttg ccggctgaaa gcgctatttc ttccagaatt gccatgattt
     1981 tttccccacg ggaggcgtca ctggctcccg tgttgtcggc agctttgatt cgataagcag
     2041 catcgcctgt ttcaggctgt ctatgtgtga ctgttgagct gtaacaagtt gtctcaggtg
     2101 ttcaatttca tgttctagtt gctttgtttt actggtttca cctgttctat taggtgttac
     2161 atgctgttca tctgttacat tgtcgatctg ttcatggtga acagctttga atgcaccaaa
     2221 aactcgtaaa agctctgatg tatctatctt ttttacaccg ttttcatctg tgcatatgga
     2281 cagttttccc tttgatatgt aacggtgaac agttgttcta cttttgtttg ttagtcttga
     2341 tgcttcactg atagatacaa gagccataag aacctcagat ccttccgtat ttagccagta
     2401 tgttctctag tgtggttcgt tgtttttgcg tgagccatga gaacgaacca ttgagatcat
     2461 acttactttg catgtcactc aaaaattttg cctcaaaact ggtgagctga atttttgcag
     2521 ttaaagcatc gtgtagtgtt tttcttagtc cgttatgtag gtaggaatct gatgtaatgg
     2581 ttgttggtat tttgtcacca ttcattttta tctggttgtt ctcaagttcg gttacgagat
     2641 ccatttgtct atctagttca acttggaaaa tcaacgtatc agtcgggcgg cctcgcttat
     2701 caaccaccaa tttcatattg ctgtaagtgt ttaaatcttt acttattggt ttcaaaaccc
     2761 attggttaag ccttttaaac tcatggtagt tattttcaag cattaacatg aacttaaatt
     2821 catcaaggct aatctctata tttgccttgt gagttttctt ttgtgttagt tcttttaata
     2881 accactcata aatcctcata gagtatttgt tttcaaaaga cttaacatgt tccagattat
     2941 attttatgaa tttttttaac tggaaaagat aaggcaatat ctcttcacta aaaactaatt
     3001 ctaatttttc gcttgagaac ttggcatagt ttgtccactg gaaaatctca aagcctttaa
     3061 ccaaaggatt cctgatttcc acagttctcg tcatcagctc tctggttgct ttagctaata
     3121 caccataagc attttcccta ctgatgttca tcatctgagc gtattggtta taagtgaacg
     3181 ataccgtccg ttctttcctt gtagggtttt caatcgtggg gttgagtagt gccacacagc
     3241 ataaaattag cttggtttca tgctccgtta agtcatagcg actaatcgct agttcatttg
     3301 ctttgaaaac aactaattca gacatacatc tcaattggtc taggtgattt taatcactat
     3361 accaattgag atgggctagt caatgataat tactagtcct tttcctttga gttgtgggta
     3421 tctgtaaatt ctgctagacc tttgctggaa aacttgtaaa ttctgctaga ccctctgtaa
     3481 attccgctag acctttgtgt gttttttttg tttatattca agtggttata atttatagaa
     3541 taaagaaaga ataaaaaaag ataaaaagaa tagatcccag ccctgtgtat aactcactac
     3601 tttagtcagt tccgcagtat tacaaaagga tgtcgcaaac gctgtttgct cctctacaaa
     3661 acagacctta aaaccctaaa ggcttaagta gcaccctcgc aagctcgggc aaatcgctga
     3721 atattccttt tgtctccgac catcaggcac ctgagtcgct gtctttttcg tgacattcag
     3781 ttcgctgcgc tcacggctct ggcagtgaat gggggtaaat ggcactacag gcgcctttta
     3841 tggattcatg caaggaaact acccataata caagaaaagc ccgtcacggg cttctcaggg
     3901 cgttttatgg cgggtctgct atgtggtgct atctgacttt ttgctgttca gcagttcctg
     3961 ccctctgatt ttccagtctg accacttcgg attatcccgt gacaggtcat tcagactggc
     4021 taatgcaccc agtaaggcag cggtatcatc aacaggctta cccgtcttac tgtcgggaat
     4081 tcaattgata tttgcttgct cttccccata ttttcctgct taccgccatt cagctggtag
     4141 tacctgtcgc aaattcttta cagtttttaa actaatgaga cgaatctgat cgacgcaaaa
     4201 agtccgtatg cctactatta gctcacggtt ataaactcga ggcataatgt gcctgtcaaa
     4261 tggacgaagc agggattctg caaaccctat gctactccgt caagccgtca attgtctgat
     4321 tcgttaccaa ttatgacaac ttgacggcta catcattcac tttttcttca caaccggcac
     4381 ggaactcgct cgggctggcc ccggtgcatt ttttaaatac ccgcgagaaa tagagttgat
     4441 cgtcaaaacc aacattgcga ccgacggtgg cgataggcat ccgggtggtg ctcaaaagca
     4501 gcttcgcctg gctgatacgt tggtcctcgc gccagcttaa gacgctaatc cctaactgct
     4561 ggcggaaaag atgtgacaga cgcgacggcg acaagcaaac atgctgtgcg acgctggcga
     4621 tatcaaaatt gctgtctgcc aggtgatcgc tgatgtactg acaagcctcg cgtacccgat
     4681 tatccatcgg tggatggagc gactcgttaa tcgcttccat gcgccgcagt aacaattgct
     4741 caagcagatt tatcgccagc agctccgaat agcgcccttc cccttgcccg gcgttaatga
     4801 tttgcccaaa caggtcgctg aaatgcggct ggtgcgcttc atccgggcga aagaaccccg
     4861 tattggcaaa tattgacggc cagttaagcc attcatgcca gtaggcgcgc ggacgaaagt
     4921 aaacccactg gtgataccat tcgcgagcct ccggatgacg accgtagtga tgaatctctc
     4981 ctggcgggaa cagcaaaata tcacccggtc ggcaaacaaa ttctcgtccc tgatttttca
     5041 ccaccccctg accgcgaatg gtgagattga gaatataacc tttcattccc agcggtcggt
     5101 cgataaaaaa atcgagataa ccgttggcct caatcggcgt taaacccgcc accagatggg
     5161 cattaaacga gtatcccggc agcaggggat cattttgcgc ttcagccata cttttcatac
     5221 tcccgccatt cagagaagaa accaattgtc catattgcat cagacattgc cgtcactgcg
     5281 tcttttactg gctcttctcg ctaaccaaac cggtaacccc gcttattaaa agcattctgt
     5341 aacaaagcgg gaccaaagcc atgacaaaaa cgcgtaacaa aagtgtctat aatcacggca
     5401 gaaaagtcca cattgattat ttgcacggcg tcacactttg ctatgccata gcatttttat
     5461 ccataagatt agcggatcct acctgacgct ttttatcgca actctctact gtttctccat
     5521 acccgttttt ttgggctagc gaattcgagc tcggtacccg ggatgtttga agaaactatt
     5581 aaaaaacaat ttgaattgtt agacataagt aacttcaatg ttgatatatc tcatcgtctc
     5641 cttttcgtgt gtggtggcaa agtcgatgta cgtgcaccaa ttccccccag ttttagggat
     5701 agactactca cttacacagc aaaaaatgcc tctgaacttc acgaacattt cattctagct
     5761 gagacattca aggattattt taaagagaat gcctatccgg accttctggt gtttgaagat
     5821 gatatagcca gtatttcatc tcttattatc atttttttag aaagcccagg atcattagtt
     5881 gagttaggta tattttgcaa caaatcagaa ctatttaaaa aaatactgat tgttgcttca
     5941 gcggaagaag tatacggtga ggactcattc atttatttag gcccactaga gtatattaag
     6001 aagaaagtct catcctctgt tgtaatctat ccttggccgg atcctgaagt cttaaaatac
     6061 gataatgatt ttcttgatga tttatgtgta aatataaaag aaaaactttc atcaatccca
     6121 aaaacagaac aattcagtaa ggataattct ggacatatag cacttctaat tactgaaatt
     6181 atatctcttt gcgccccaat acaacttagt gaaatagaat cagccctaaa ctcattaggg
     6241 attaatattt caacaaaaat tattaacaga tcaatttatt tacttcagaa agttggtttt
     6301 atagacgttt tatcatatag ctcaaacaaa tattactttc cattaaaaga gcgcaaatgg
     6361 gtaaaatttg gtaaaactaa agataataaa ctcatagata atcaacaatt aaaaatgaag
     6421 gttagacaat cttttgtaac attaaccgat ccattatcaa aacgaagaat tacagcgctg
     6481 cgtcaaataa tagcaaaaaa agaaatggct gaggagatta attaa
//
