LOCUS       sequence-448698-        6517 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of ECOR31 Hachiman HamB (∆HamA) under the pTet promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6517)
  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 6517)
  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: pBA1464_pTet-ECOR31Ham_DELhamA_Mut
FEATURES             Location/Qualifiers
     source          1..6517
                     /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(4801..4835)
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     CDS             complement(4906..5565)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     5480..5499
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     primer_bind     complement(5804..5821)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5938..6483)
                     /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(6018..6037)
                     /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 ctgatggaaa aggtgattag cgatgccagc
     1261 aacagctgat gaaattatcg aagctattaa ggaagcttca gctgtaggtt ttcgcgggcg
     1321 cctgattgca cgcgggcaag cccggtcggt gatctggcga gacggtgact tgccgccgga
     1381 tgcaccggag tttagcgcac tgctcagtca ggaccttcaa ggctatgctt atgccctaat
     1441 cgatctcggc cttcgtcttc gagaacttaa tggagacgac gcctatgcgc gaattgcttt
     1501 tgaacaggcc ggaaccgcac tcgagtcggc tattgctaaa ggaaaacgcg acagccggga
     1561 tacagatttt cacttcgtaa tggctgctgc aagctatcac cttgcgcatc tttcagcacg
     1621 agcctattcg ttactcgcaa tggttggtca ggatgataat ttttcaccaa tagagagggc
     1681 tcttacacaa ctgatccggc gtgatttgcg tactctgcgc gataatgcgt tgggctttcg
     1741 gctcagaggt gatggcagtg acgtgaaaat tactgaaatc ctccaggctc gcctaaactt
     1801 gccgcaagac gagaacggtg acagtgagtc agaggaggac atcctttttg atgggcttga
     1861 tttggcactg actgacgcct atatgtcggc catttctctc taccttctcg ctgtagaaag
     1921 gggagaatca cgtttgcttt ccagggctat tgaaaaatta agaattagtc taagtatatg
     1981 tgctcagttt aatatgctac cccagtggtg gctaaacttt attaccattc atctgttatc
     2041 tgacctttgg tcagatacat tccatgaaag attgccactg gttccagttg gtggtgacgc
     2101 tgctgaatgg cccgcgcttc gggaactgtt tattgcactg cttcaaagac gacctcgagc
     2161 cgaaattgat ctctggccat cgcagagaga ggccgccgga cgctctgtta atgacaatga
     2221 cgatttggtt gtttcgttgc caaccagcgc tggaaaaact cgtattgctg aattgtgtat
     2281 tcttcgctgt ctggcgggtg gcaagcgtgt tgttttcatc accccgcttc gtgcgctttc
     2341 tgcacaaaca gaagccactc tctccagaac atttgggccg cttgggaaaa caatatcaat
     2401 gctgtatggc agcattggcg taagtggtat ggatgaggat gcaattcgcc agcgtgacat
     2461 cgtcgtcgcc acgcctgaaa aacttgactt tgctctccgg aatgacccgt caattattaa
     2521 tgatgtagga ctttttattt ttgatgaggg gcacatgatt ggggccgatg aacgagaggt
     2581 tcggtatgaa gttcaaattc aacggctgtt gcgcagacag gatgccgata cgcgcagaat
     2641 tgtctgcctc tcggcaattc tacctgatgg tgagcaactc gatgattttg ccggttggct
     2701 ccgtcgtgac aagcccggag gaccaatcaa aaataattgg cgaccgaccc gcttgcagtt
     2761 tggcgaggtc atatggtccg ctccagcagg tcggcttaat ctcagcgtcg gttatgaggc
     2821 ggcttgggta tcgcgtttta tcgtcagcag acaaccgcca aaggttaaat tacctaataa
     2881 aaagcagcga acaaaaatgt ttccctctga taacaaggag ctctgcttag cgacagcctg
     2941 gcgtctaatt gaagatggac agaccgtact tatctactgc ccacttcgcc ggagcgtcga
     3001 accgtttgcc gagacaattg tcgatctgca tcagcgcgga ctactgccat ctttatttga
     3061 tgctgccccg gacattctgg atacggctat ctcgctggga gaagaatggt tgggggcaca
     3121 cagcccgata cttgcctgcc tacggttggg cgtggctctt catcatggcg cattgcctac
     3181 ggcttaccgc aaagaaatcg aacgtttgct gcgagatggg gtacttaaag tgacgatctc
     3241 atcaccgacg ctagcacaag gtcttaatct ttcggcaaca gctatcgtca tgcattctct
     3301 tcatcggaat cgcgagctta tcaaagtgtc cgagttcaga aacgtcatag gcagagcagg
     3361 gcgcgcatat gtagatgttg aagggctggt gatttatcca atattcgata aagtaaataa
     3421 acgccagact aactggcata ccctcacatc tgacacgggg gcccgtgaga tggagagcgg
     3481 actaatacag cttgtctgtg tgttattgat tcgcatgcat acacggcttg gtggggattt
     3541 aaaagcactc actgaatatg tgaccaacaa cgccgtagcg tgggaatttc cagaaattat
     3601 gaccgaatct ccgcaagaaa gagatattgc gcaggccatc tgggaaaaac aactttcaac
     3661 gcttgatact gctattctga gccttcttgg tgagaatgat atcccggatg accagattga
     3721 aactgctctt gatgacattc ttcagtcttc actttggcaa cgcagtttgc agcgttatag
     3781 ggatgagaat gaacggattt tgctgaaatc gggactcctg tcccgcagcc gctacatctg
     3841 gcagcgctca acggcagcag ggcgccgggg gtatttcctg tcgggggtag gacttacaac
     3901 cgggctacga cttgatgcta ttgccgctaa ggccaaccaa ctattgattg atgcgaatgc
     3961 ggctataatg ggcggtgatg ccgaagaggc aattgctgct atcacggcgc tcgctgaaga
     4021 ggtctttacc ttctatccgt ttatacccga tccattgcct ggcgactgga gaggtatttt
     4081 acgttcctgg ctattgggtg aacccatgac taacgtcgcg aatacgcagg cgtctgaaac
     4141 gcttcaattc gtcgagaatg ggctggtgta tcgacttccg tgggcaatgg aggctattag
     4201 ggtacgtgca acagctaatg gagacctcat aggtgataca gataccacgc tagacgatta
     4261 tgagttaggt tttgcggttg ctgctgttga aacaggtacc ttgagccgtt cgtcttcact
     4321 gcttattcag gcgggtttta gctctaggct tgcagccata aaagtggtta cagacactac
     4381 tgccgatttt caatccggcc aggagcttag aaggtggctg aactctgaag aagtgataag
     4441 ccacacggat aatcatgact ggcctacacc tgaaaccaga gtgatgtggc tggaattctt
     4501 gggtagtctc agccccaaag gcagccaggt ttggagtcgt catcgttata acggaatggt
     4561 ggactggaga gatactccgg cagtgatcgg cacaccattg cagctttaca ctgttgatgg
     4621 tattcatcac gtactggctg atgatggcac tcctctggga agtataaatg gcagaattaa
     4681 cacaaacaga agaggtcttc ttagagttga ggtggatgat gagaatggtc gtgccatgtt
     4741 cgattattta ggaccggacg actttatttc tacctgaagg aggactttgg caggtgacca
     4801 aaaaaaaacg cccggcggca accgagcgtt ctgaacaaat ccagatggag ttctgaggtc
     4861 attactggat ctatcaacag gagtccaagc gagctcgata tcaaattacg ccccgccctg
     4921 ccactcatcg cagtactgtt gtaattcatt aagcattctg ccgacatgga agccatcaca
     4981 aacggcatga tgaacctgaa tcgccagcgg catcagcacc ttgtcgcctt gcgtataata
     5041 tttgcccatg gtgaaaacgg gggcgaagaa gttgtccata ttggccacgt ttaaatcaaa
     5101 actggtgaaa ctcacccagg gattggctga aacgaaaaac atattctcaa taaacccttt
     5161 agggaaatag gccaggtttt caccgtaaca cgccacatct tgcgaatata tgtgtagaaa
     5221 ctgccggaaa tcgtcgtggt attcactcca gagcgatgaa aacgtttcag tttgctcatg
     5281 gaaaacggtg taacaagggt gaacactatc ccatatcacc agctcaccgt ctttcattgc
     5341 catacgaaat tccggatgag cattcatcag gcgggcaaga atgtgaataa aggccggata
     5401 aaacttgtgc ttatttttct ttacggtctt taaaaaggcc gtaatatcca gctgaacggt
     5461 ctggttatag gtacattgag caactgactg aaatgcctca aaatgttctt tacgatgcca
     5521 ttgggatata tcaacggtgg tatatccagt gatttttttc tccattttag cttccttagc
     5581 tcctgaaaat ctcgataact caaaaaatac gcccggtagt gatcttattt cattatggtg
     5641 aaagttggaa cctcttacgt gcccgatcaa gcgctagcgg agtgtatact ggcttactat
     5701 gttggcactg atgagggtgt cagtgaagtg cttcatgtgg caggagaaaa aaggctgcac
     5761 cggtgcgtca gcagaatatg tgatacagga tatattccgc ttcctcgctc actgactcgc
     5821 tacgctcggt cgttcgactg cggcgagcgg aaatggctta cgaacggggc ggagatttcc
     5881 tggaagatgc caggaagata cttaacaggg aagtgagagg gccgcggcaa agccgttttt
     5941 ccataggctc cgcccccctg acaagcatca cgaaatctga cgctcaaatc agtggtggcg
     6001 aaacccgaca ggactataaa gataccaggc gtttccccct ggcggctccc tcgtgcgctc
     6061 tcctgttcct gcctttcggt ttaccggtgt cattccgctg ttatggccgc gtttgtctca
     6121 ttccacgcct gacactcagt tccgggtagg cagttcgctc caagctggac tgtatgcacg
     6181 aaccccccgt tcagtccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     6241 cggaaagaca tgcaaaagca ccactggcag cagccactgg taattgattt agaggagtta
     6301 gtcttgaagt catgcgccgg ttaaggctaa actgaaagga caagttttgg tgactgcgct
     6361 cctccaagcc agttacctcg gttcaaagag ttggtagctc agagaacctt cgaaaaaccg
     6421 ccctgcaagg cggttttttc gttttcagag caagagatta cgcgcagacc aaaacgatct
     6481 caagaagatc atcttattaa tcagataaaa tatttct
//
