LOCUS       sequence-500763-        6874 bp    DNA     circular SYN 23-FEB-2026
DEFINITION  Hachiman from P. fluorescens under pTet control with the dHamA 
            domain removed.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6874)
  AUTHORS   Doudna J et al.
  TITLE     Recurrent acquisition of nuclease-protease pairs in antiviral 
            immunity
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6874)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Feb 23, 2026 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pTet_PflHamMA(deltaA)B
FEATURES             Location/Qualifiers
     source          1..6874
                     /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(5158..5192)
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     CDS             complement(5263..5922)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     5837..5856
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     primer_bind     complement(6161..6178)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6295..6840)
                     /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(6375..6394)
                     /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 agagttattt taccactccc tatcagtgat agagaaaaga
     1201 attcaaaaga tcttttaaga aggagatata ctgatgaaaa tttccatgca cgatgccgca
     1261 tacggtgatt caattctcat cagcggcgcc tcgaccaaaa ttttagtgga tggtggaact
     1321 gcgaagtcct tcgctgactg gtcgccttcg ttagagggtg tcgatacgct agatgccctg
     1381 atagttactc atattgataa cgaccatatt ggcggagcaa tcaagctgct cacatcaaag
     1441 cacgcaacta aagtaaaaga agtttatttc aatggcatta accaaatcct agatgtggag
     1501 gattgtgata ttggtgatcg tagtgacgat ctagaattat ccgcaatcat agcaaacacc
     1561 acgtctagag acgaagacca gttgataggg tattctgaag gcacatcgtt atccttaata
     1621 ttacagcagc aaaatatccc cttaaataaa gcaaccgaag gcaggcaaat ccacagaacg
     1681 agccttaata aaaacataat aataggagaa tttgaaattg aactgatagg cccaagcaga
     1741 aagagcttgt taaaaatcca acaaaaatgg cttgaactat tagaaagtaa agaaatcaaa
     1801 agacgcctta tcagcccaag atatagcgtt gcattcgaaa gctttatcag ctcgattgat
     1861 acctcagtaa agcatgatga ggaaatttca agtctaagtg aaaacataga gcatttggca
     1921 aatcaaccat ttactaatga cacatcaccc ccaaacaaaa gcagcatctc tttccttcta
     1981 aaaactgaaa acaaaagcat tttaatgctc ggagattccg acgcagagac tatagaggca
     2041 tggttgaact cgaagtctgc agatagcatc acagttgatg cggtaaagct accccatcac
     2101 ggaagccgaa ataactttag tgctaactta gtacgcagaa tacgcactgc gaactattta
     2161 atttctacta atggcgataa attttcacac cctaacattg agaccttagc tagaatcatt
     2221 aagttttctc ctgaccgcga ggcaaaaata ctactaaacc acgaaatccc ccacctcaca
     2281 aactccttcc tgagcaacgc caacagttat gaaggaaata tcagcatcca ccagggcata
     2341 aaggaatttg agctttgagt aatgcactga ctgacgagta caaactcgat aacgtcttta
     2401 tcctagaagc ctatgacgac ctaggtgaac ccatttcaac aggcagttgc ttttcggtgg
     2461 gtgcgtctat tttttttaca tcttggcacg tcgtcaaaaa tgcctacacg cttcggattt
     2521 tccaaagtag agatgattat tttcgtcaag agttccatgc cgctaaatgt gttttttacg
     2581 ataaagaaac tgacttcgca atactgtcat cagacagcct tacgatcaaa tcaaaaattg
     2641 aaataggaat atacaaagct gagctgaatg atgaggtaaa agctataggg tacgcggccg
     2701 aaaaaagcag attgcatttc cctataacta gtaaaatatc aaacgtcttt gatttctcta
     2761 caacatataa atatgacttt gagtttggaa agccggctga gctagacaaa gcaaatggct
     2821 tatccggggg gccagtattc tttaaaggct acgctattgg cttgcttgca ctgcaaactg
     2881 tgggatcggc attgcaagca gtatctttcc atacaatatt tgaacggcca tccgctcttg
     2941 aagccattga aagccaaggg ttttgaatac ggaaatttat aacaaccttc gtaagcaaat
     3001 cgacaactct gctttcaccg gatggcaaag gttattcgtt gaacttataa atctatacaa
     3061 cttacatgat gacaatcaat tcttgattca ggacctgacg ctgagatgcc tagagaggcg
     3121 tggggaattc aaacagcttc atggtctgtt gaacatattg atttccgagc taggtttgtt
     3181 cccgtactta gatccagatc tgatgacgtc gaaagacagc ataaggatga atatttttaa
     3241 gtcgcctata gctggcgata taacattcca cctaaaacag gctgagattt attacaggat
     3301 tgtaaatgga gataacatcg tactcagtgc acctacaagc ttcggaaaaa gcttagttat
     3361 agatgctctc atcagctctg aagaatacga caacattgtg gtagtagtcc caaccatagc
     3421 tttgatggat gagttacgaa agaaatttag caaatataaa gaaaactaca agataatcac
     3481 tcagtcctct caagagccag cgacaaaaaa tctatatatc tatactcaag aaagggtctt
     3541 ggaatgcaag aaccttgaac gagtagactt ctttattatc gatgagtttt acaaattagc
     3601 acctctcaat acatctgact atcgaagcga cagactaaat ctggcatttc acaaactttt
     3661 taaaatgtgc agtcgctttt acatgctcgg ccccaacatt aatggcctgg ccgaaggcat
     3721 agaagacaaa ctaaattgca attttgtaaa atatgacaat tacaaaactg tagtagctaa
     3781 cgaatatcgc tacattatcc gtgaagatgg gaatgataaa gaaaaagaca ttgatcgcga
     3841 cgcgcactta gtcaatattt tgcaaaaaat cggtcaaaac gaacagactt taatttactg
     3901 caagactcca aaacgagcag cctcgctagt aaaaaagatc atgaaactca gagagaacca
     3961 agagataaga aaaggagagt tttctgactg gctgagagag acatatcacg atgactggtc
     4021 gatggcagac gcagtggaac gcgggattgc ctgtcaccat ggcagacttc ctcgtgtcgt
     4081 ctccgggtat gtggtggaag cgttcaatga aaagctcata aatattttgg tttgcacctc
     4141 aactttgatt gagggtgtta acacaaatgc tagaagcgtc ataatttacg atgactgtat
     4201 tcgagggacc acccaactgg attcgttcac ttttaacaat atctctggac gtagtgggcg
     4261 tatgtttgaa cactttgttg gagatataca tgttattgga cgaaaaccat cagcagagct
     4321 tccttggatc gacgtcccaa tagtgactca agatgactcc gcctctagac ccatgctgct
     4381 aagcataatc gacgagctaa gcaccgaaca aagggctagt ttatcgagat atacaactca
     4441 acaatatgta cctatcactt tacttaataa gcatctcggt gttcctccgg aaaggataat
     4501 aaaattcgca gaagccttaa tggagaaatg tggcgcctgg aactcaatta tggcatggac
     4561 gggaaaattc ccaactagcg agcaactaag acacctatgc aacatcatgt tcgatcactt
     4621 cgcaatatca cgaatgggcg gaggatgcgt caaatcaaaa aagcagctgt tttatagagt
     4681 gctcaacatt gtaaatagaa aagaagacaa agatctaatt caaagcgaat atgatttttg
     4741 gtccccacga aacgcggaat atgagattga tgacgcagtg caaagcgttt tcgatttcaa
     4801 aagaaactta attggctaca acctccctaa gattattttg gcgataaacg acgtacagcg
     4861 cttaatattt tctcgattca attaccgttg tggcgactac gcagcttttg cccactctct
     4921 agaagcaatg tttttaccac cttcaattgt ggctcttgaa gagtttggaa taccaactcg
     4981 tgtagcacaa aaaatatacc aaattagcaa cgtcggcgag gacgccaccg tagatttaat
     5041 cctagagcag gtaagatcgc acccgggtct ctacatatcg aaatttgatg attttgaaaa
     5101 aactctcctg aaccgagcaa ttcaatatct ttaaaggagg actttggcag gtgaccaata
     5161 aaaaacgccc ggcggcaacc gagcgttctg aacaaatcca gatggagttc tgaggtcatt
     5221 actggatcta tcaacaggag tccaagcgag ctcgatatca aattacgccc cgccctgcca
     5281 ctcatcgcag tactgttgta attcattaag cattctgccg acatggaagc catcacaaac
     5341 ggcatgatga acctgaatcg ccagcggcat cagcaccttg tcgccttgcg tataatattt
     5401 gcccatggtg aaaacggggg cgaagaagtt gtccatattg gccacgttta aatcaaaact
     5461 ggtgaaactc acccagggat tggctgaaac gaaaaacata ttctcaataa accctttagg
     5521 gaaataggcc aggttttcac cgtaacacgc cacatcttgc gaatatatgt gtagaaactg
     5581 ccggaaatcg tcgtggtatt cactccagag cgatgaaaac gtttcagttt gctcatggaa
     5641 aacggtgtaa caagggtgaa cactatccca tatcaccagc tcaccgtctt tcattgccat
     5701 acgaaattcc ggatgagcat tcatcaggcg ggcaagaatg tgaataaagg ccggataaaa
     5761 cttgtgctta tttttcttta cggtctttaa aaaggccgta atatccagct gaacggtctg
     5821 gttataggta cattgagcaa ctgactgaaa tgcctcaaaa tgttctttac gatgccattg
     5881 ggatatatca acggtggtat atccagtgat ttttttctcc attttagctt ccttagctcc
     5941 tgaaaatctc gataactcaa aaaatacgcc cggtagtgat cttatttcat tatggtgaaa
     6001 gttggaacct cttacgtgcc cgatcaagcg ctagcggagt gtatactggc ttactatgtt
     6061 ggcactgatg agggtgtcag tgaagtgctt catgtggcag gagaaaaaag gctgcaccgg
     6121 tgcgtcagca gaatatgtga tacaggatat attccgcttc ctcgctcact gactcgctac
     6181 gctcggtcgt tcgactgcgg cgagcggaaa tggcttacga acggggcgga gatttcctgg
     6241 aagatgccag gaagatactt aacagggaag tgagagggcc gcggcaaagc cgtttttcca
     6301 taggctccgc ccccctgaca agcatcacga aatctgacgc tcaaatcagt ggtggcgaaa
     6361 cccgacagga ctataaagat accaggcgtt tccccctggc ggctccctcg tgcgctctcc
     6421 tgttcctgcc tttcggttta ccggtgtcat tccgctgtta tggccgcgtt tgtctcattc
     6481 cacgcctgac actcagttcc gggtaggcag ttcgctccaa gctggactgt atgcacgaac
     6541 cccccgttca gtccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     6601 aaagacatgc aaaagcacca ctggcagcag ccactggtaa ttgatttaga ggagttagtc
     6661 ttgaagtcat gcgccggtta aggctaaact gaaaggacaa gttttggtga ctgcgctcct
     6721 ccaagccagt tacctcggtt caaagagttg gtagctcaga gaaccttcga aaaaccgccc
     6781 tgcaaggcgg ttttttcgtt ttcagagcaa gagattacgc gcagaccaaa acgatctcaa
     6841 gaagatcatc ttattaatca gataaaatat ttct
//
