LOCUS       sequence-444931-        3757 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of ECOR31 Hachiman HamA (∆HamB) under the pTet promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3757)
  AUTHORS   Tuck OT, Adler BA, Armbruster EG, Lahiri A, Hu JJ, Zhou J, Pogliano 
            J, Doudna JA
  TITLE     Hachiman is a genome integrity sensor.
  JOURNAL   bioRxiv [Preprint]. 2024 Feb 29:2024.02.29.582594. doi: 
            10.1101/2024.02.29.582594.
  PUBMED    38464307
REFERENCE   2  (bases 1 to 3757)
  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: "bioRxiv 
            [Preprint]. 2024 Feb 29:2024.02.29.582594. doi: 
            10.1101/2024.02.29.582594."
COMMENT     Sequence Label: pBA1465_pTet-ECOR31Ham_DELhamB_Mut
FEATURES             Location/Qualifiers
     source          1..3757
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             133..162
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             178..195
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(251..268)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      421..467
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(499..1125)
                     /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     1030..1049
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     promoter        1141..1196
                     /gene="tetR"
                     /label=tetR/tetA promoters
                     /note="overlapping promoters for bacterial tetR and tetA"
     protein_bind    1177..1195
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     terminator      complement(2041..2075)
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     CDS             complement(2146..2805)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     2720..2739
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     primer_bind     complement(3044..3061)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3178..3723)
                     /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(3258..3277)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 agatttcagt gcaatttatc tcttcaaatg tagcacctga agtcagcccc atacgatata
       61 agttgtcacc tgcccgaacc agagacttat cctgaactgg ccgataattg cagacgaacg
      121 atgcttgggc ccgaacaaaa actcatctca gaagaggatc tgaatagcgc cgtcgaccat
      181 catcatcatc atcattgagt ttaaacgctc tccagcttgg gtgttttggc ggatgagaga
      241 agattttcag cctgatacag attaaatcag aacgcagaag cggtctgata aaacagaatt
      301 tgcctggcgg cagtagcgcg gtggtcccac ctgaccccat gccgaactca gaagtgaaac
      361 gccgtagcgc cgatggtagt gtggggtctg cccatgcgag agtaggcaac tgccaggcat
      421 caaataaaac gaaaggctca gtcgaaagac tgggcctttc gttttatctg ttgtttgtcg
      481 gtgaacttca gcgacgtctt aagacccact ttcacattta agttgttttt ctaatccgca
      541 tatgatcaat tcaaggccga ataagaaggc tggctctgca ccttggtgat caaataattc
      601 gatagcttgt cgtaataatg gcggcatact atcagtagta ggtgtttccc tttcttcttt
      661 agcgacttga tgctcttgat cttccaatac gcaacctaaa gtaaaatgcc ccacagcgct
      721 gagtgcatat aatgcattct ctagtgaaaa accttgttgg cataaaaagg ctaattgatt
      781 ttcgagagtt tcatactgtt tttctgtagg ccgtgtacct aaatgtactt ttgctccatc
      841 gcgatgactt agtaaagcac atctaaaact tttagcgtta ttacgtaaaa aatcttgcca
      901 gctttcccct tctaaagggc aaaagtgagt atggtgccta tctaacatct caatggctaa
      961 ggcgtcgagc aaagcccgct tattttttac atgccaatac aatgtaggct gctctacacc
     1021 tagcttctgg gcgagtttac gggttgttaa accttcgatt ccgacctcat taagcagctc
     1081 taatgcgctg ttaatcactt tacttttatc taatctagac atcattaatt cctaattttt
     1141 gttgacactc tatcgttgat atagttattt taccactccc tatcagtgat agagaaaaga
     1201 attcaaaaga tcttttaaga aggagatata ctgatggcaa atttcgagga ctggtgtgat
     1261 tcgacagagc gtaatatcag cgaccattat ttgcagtcaa taacggccag ggatgccgag
     1321 tgtatgttcg gcgttcaggt tatggccgcg ctaatacctg aacactatgc gtcaccaaga
     1381 aatatcgcta acgcatttga agctctaggt aaaccggggc tggcggcata cattgctgga
     1441 aaattgcctg aaacgaaaca gattcgttcc ggtgatttgg gtgagatatt tgcaactgaa
     1501 tggataaatg cacgtagcaa tggctataaa acgcctataa aaagattgcg ctggaaagat
     1561 catcgcaata tgtcgatgcg tggtgaggac gtcattggta tttacataga tcagtcttct
     1621 caacaacttt ttttccttaa aaccgaagcg aaaagtcgcg ccaaaatgac gggagaggtg
     1681 gtttctgaag cccgcgataa cttgaataaa gagcaagggc tgccatcttc acatgcccta
     1741 atgttcatag cagatcgcct gaatgaacag ggtgaggagc ttttggctaa agcgatctta
     1801 aatgcaacgc ttcggcaagg aattgttcca ggatgcgtcc ggcatcttat ctttctatta
     1861 tcgggcaact cttcagaaac tatgctgacc acttcaattg aaaagtatac aggtcaaaat
     1921 aatcagtggg gtgtttgtct aagaattgct cgccatggtg agtttattgc cgcaactttt
     1981 gaaaaggtga ttagcgatgc cagcaacagc tgattgaagg aggactttgg caggtgacca
     2041 aaaaaaaacg cccggcggca accgagcgtt ctgaacaaat ccagatggag ttctgaggtc
     2101 attactggat ctatcaacag gagtccaagc gagctcgata tcaaattacg ccccgccctg
     2161 ccactcatcg cagtactgtt gtaattcatt aagcattctg ccgacatgga agccatcaca
     2221 aacggcatga tgaacctgaa tcgccagcgg catcagcacc ttgtcgcctt gcgtataata
     2281 tttgcccatg gtgaaaacgg gggcgaagaa gttgtccata ttggccacgt ttaaatcaaa
     2341 actggtgaaa ctcacccagg gattggctga aacgaaaaac atattctcaa taaacccttt
     2401 agggaaatag gccaggtttt caccgtaaca cgccacatct tgcgaatata tgtgtagaaa
     2461 ctgccggaaa tcgtcgtggt attcactcca gagcgatgaa aacgtttcag tttgctcatg
     2521 gaaaacggtg taacaagggt gaacactatc ccatatcacc agctcaccgt ctttcattgc
     2581 catacgaaat tccggatgag cattcatcag gcgggcaaga atgtgaataa aggccggata
     2641 aaacttgtgc ttatttttct ttacggtctt taaaaaggcc gtaatatcca gctgaacggt
     2701 ctggttatag gtacattgag caactgactg aaatgcctca aaatgttctt tacgatgcca
     2761 ttgggatata tcaacggtgg tatatccagt gatttttttc tccattttag cttccttagc
     2821 tcctgaaaat ctcgataact caaaaaatac gcccggtagt gatcttattt cattatggtg
     2881 aaagttggaa cctcttacgt gcccgatcaa gcgctagcgg agtgtatact ggcttactat
     2941 gttggcactg atgagggtgt cagtgaagtg cttcatgtgg caggagaaaa aaggctgcac
     3001 cggtgcgtca gcagaatatg tgatacagga tatattccgc ttcctcgctc actgactcgc
     3061 tacgctcggt cgttcgactg cggcgagcgg aaatggctta cgaacggggc ggagatttcc
     3121 tggaagatgc caggaagata cttaacaggg aagtgagagg gccgcggcaa agccgttttt
     3181 ccataggctc cgcccccctg acaagcatca cgaaatctga cgctcaaatc agtggtggcg
     3241 aaacccgaca ggactataaa gataccaggc gtttccccct ggcggctccc tcgtgcgctc
     3301 tcctgttcct gcctttcggt ttaccggtgt cattccgctg ttatggccgc gtttgtctca
     3361 ttccacgcct gacactcagt tccgggtagg cagttcgctc caagctggac tgtatgcacg
     3421 aaccccccgt tcagtccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     3481 cggaaagaca tgcaaaagca ccactggcag cagccactgg taattgattt agaggagtta
     3541 gtcttgaagt catgcgccgg ttaaggctaa actgaaagga caagttttgg tgactgcgct
     3601 cctccaagcc agttacctcg gttcaaagag ttggtagctc agagaacctt cgaaaaaccg
     3661 ccctgcaagg cggttttttc gttttcagag caagagatta cgcgcagacc aaaacgatct
     3721 caagaagatc atcttattaa tcagataaaa tatttct
//
