LOCUS       sequence-416251-        7289 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  JAGBLH010000220 Cas8-HNH full system (E. coli codon optimized) in 
            pACYCDuet-1 with Lac promoters.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7289)
  AUTHORS   Altae-Tran H, Kannan S, Suberski AJ, Mears KS, Demircioglu FE, 
            Moeller L, Kocalar S, Oshiro R, Makarova KS, Macrae RK, Koonin EV, 
            Zhang F
  TITLE     Uncovering the functional diversity of rare CRISPR-Cas systems with 
            deep terascale clustering.
  JOURNAL   Science. 2023 Nov 24;382(6673):eadi1910. doi: 
            10.1126/science.adi1910. Epub 2023 Nov 23.
  PUBMED    37995242
REFERENCE   2  (bases 1 to 7289)
  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: "Science."; 
            date: "2023-11-24"; volume: "382(6673):eadi1910. doi"; pages: " 
            10.1126/science.adi1910. Epub 2023 Nov 23"
COMMENT     Sequence Label: pHS1801
FEATURES             Location/Qualifiers
     source          1..7289
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(12..83)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     promoter        complement(222..256)
                     /label=J23119 promoter
                     /note="bacterial promoter (Registry of Standard Biological 
                     Parts BBa_J23119)"
     promoter        join(307..312,313..330,331..337)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    340..364
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     CDS             1101..1115
                     /codon_start=1
                     /product="enterokinase recognition and cleavage site"
                     /label=enterokinase site
                     /translation="DDDDK"
     primer_bind     complement(3787..3805)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3801..3848
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(4069..4728)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     4643..4662
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(4729..4831)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(5223..5240)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5357..5902)
                     /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(5437..5456)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(6127..7209)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=lacI
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     7171..7190
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(7210..7287)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 ggatctcgac ggagagcgtt caccgacaaa caacagataa aacgaaaggc ccagtctttc
       61 gactgagcct ttcgttttat ttgggcgcgc caaaaaaatc gacctgcagg cgcgccgagc
      121 tcgaattcgg atccttctaa accgcctatc cggcggtaca ctttttaaca gtggccttat
      181 taaatgactt ctccttctaa accgcctatc cggcggtaca cgctagtatt atacctagga
      241 ctgagctagc tgtcaacatt aattcctaat ttttctctcc cttatgcgac tcctgcatta
      301 ggaaatttta cactttatgc ttccggctcg tatgttgggg gaattgtgag cggataacaa
      361 ttcccctgta gaaataattt tgtttaactt taataaggag atataccatg gcagctaata
      421 aaaaggcaac caatgtcacg ctaaaatccc gccccgaaaa tctatcattt gctaggtgtt
      481 taaacacgac tgaggcgaaa ttctggcaga ccgattttct gaaacgccat acgttcaaat
      541 taccgctgct gattaccgat aaggcggtcc tcgcgagcaa aggccacgaa atgccgcctg
      601 ataagctgga aaaagagatc atggacccga atccgcaaaa gtcccagtct tgtaccctgt
      661 cgaccgagtg cgatacgctg cgtattgact ttggtattaa agtgctgccg gttaaagaaa
      721 gcatgtacag ctgcagcgat tacaactatc gtaccgcaat ttatcagaaa atcgacgaat
      781 acatcgcgga ggacggcttc ttgaccttgg cgaagcgcta tgtgaataac atcgcgaatg
      841 cccgtttttt gtggcgtaat cgtaaaggtg ctgagatcat cgaaaccatt gttaccattg
      901 aggacaagga gtacccgagc tttaatagca agtccttcaa cctggatact ttcgtggagg
      961 acaacgccac catcaacgaa atcgctcagc agattgccga tacgtttgct ggcaagcgcg
     1021 aatacttgaa tatttacgtc acctgctttg ttaagatcgg ttgtgcaatg gaagtgtatc
     1081 catcgcaaga aatgaccttt gatgacgatg acaagggtaa gaaactgttc aagttcgagg
     1141 gtagcgcagg tatgcatagc caaaagatca acaacgcgct gcgtaccatt gacacatggt
     1201 atccggacta caccacctat gaatttccga ttccggtaga gaactacggc gcggcgcgtt
     1261 caatcggcat cccgttcaga ccagacacca aatctttcta taaacttatc gaccgcatga
     1321 ttctgaaaaa cgaggatctg ccgatcgagg acaagcacta cgttatggca attttgattc
     1381 gaggcggcat gttcagcaag aaacaagaaa aataagcatt caggaggaaa cgatatgttt
     1441 agtcaaattc ttatcataaa accaggcaca ggcatatccc ccaatattat aatctcagaa
     1501 gatatatttc cagttctgca tagcttgttc gtggagcacg ataaaaagtt tggtatcacg
     1561 ttcccggcat actcctttga taaaaagggc catctgggta atatcattga agttctgagc
     1621 gaagataaag aggcgctggc gagcctgtgt ctggaggagc acttggcgga agtcaccgac
     1681 tatgtgaaag ttaagaagga gatcacgttc accgacgact acgtgttgtt taaacgcatt
     1741 agagaagaga accaatatga aaccactgcc cgtcgtatgc gtaaacgcgg tcacaccgaa
     1801 ctcggccgtc cgctggagat gcatattaag aagaagaacc agcaaatttt ctgccacgcg
     1861 tacatcaagg tgaaaagcgc atccaccggt cagagctata acatctttct tgctccgacc
     1921 gacatcaaac acggctcgtt ttccgcttat ggacttcttc gaggtgatac tcatgcttag
     1981 aaataaaata ttggctgcca tatcgcaaaa aatccctgag gaacaaaaaa taaataagta
     2041 tattgaaggg ctgtttcaaa gcattgataa gaaccacctt gctacgcatg tggccaaatt
     2101 caccgaaacc aacagcccgg gtaatatagg tgcgtatgat atcttgtcca gcgacatgaa
     2161 ctgcggttac ctggacaccg caaatgccgg ttggaaagaa ccggatatcg ttaccaacga
     2221 tgcgaaatac aaacgtcctc agggctttgt tgctatggaa atgtccgacg gccgtaccgt
     2281 gatggagcac ctacaagaag attctgcaga attaagacac gagatggaag agttgaccga
     2341 taagtacgac gagatccgtg acggtatttt gaacatgccg agcatgcagc cgtatcgcac
     2401 gaaccagttt atcaagcaag ttttcttccc ggtcggtggt agctatcatc tcctcagcat
     2461 tctcccaagc accgtactga attacgaggt gagcgaccgc ctgtatcgta gcaagatccc
     2521 gaagattcgc ctgcgcttgc tgtcgagcaa cgcggcatcg acgaccggta gccgtctggt
     2581 ttccaagaac aagtggccgc ttgttttcca agcgctgccg ccaaagttcc tggaaaaaaa
     2641 cctggctaaa gcgctggaca aggagtatct gctgccggac atcaacatcg acgagttgga
     2701 aggcgttgat aatggttgtc tgattgatga ggccctgtta ccgctgatca tcgatgaggg
     2761 caaacgcaag ggtgaaggca attatcgtcc gcgtcacctg cgtgacgaac gtaaagagga
     2821 gactgtccag gcgtttttgg acaaatacgg ctactgcaat attccggtgg gctacgaggt
     2881 gcatcatatt gtgccgctgt ctcagggcgg tgcggatagt atcaaaaaca tgattatgct
     2941 gtctatcgaa caccacgaaa gggttactga agctcatgca tcatatttca agtggagaaa
     3001 tacatgatga agggatatat actgcttgaa aaagtaaata ttgaaaacgc taatgcattt
     3061 aataacatta tagtaggaat cccggctatc acctcattcc tgggctttgc aagagcgcta
     3121 gagcgcaaac tgaacgcgaa agagatcgct atccgtatca acggtgttgg cctggaattt
     3181 catgaatatg aattaaaagg ttacaagaac aaacgtggtc agtatgtgac cagctgtccg
     3241 ctgccgggtt ccattccagg tcagaacgag aagaagctgg acgctcacat catgaatcag
     3301 gcatatattg atctgaatat gagctttttg ctggaggtgg agggtccgca tgttgatatg
     3361 tctacgtgca aatctattaa atcgaccatg gaaacgctgc gtattgcggg tggcattatt
     3421 cgcaactaca agaagatccg cttgattgac acccttgccg atatcccgta cggctacttc
     3481 ttgaccctcc gtcaggataa tctgaatgat gcggcaggcg acgacatgct ggataagatg
     3541 attcatgcgt tgcaacaaga ggacactttg gttccgattg ccgttggctt caaggcgctg
     3601 agcgaagtcg gtcacgtgga gggccaacgt gatccggaaa aggaccactg cttcgtggag
     3661 tccatcttca gcctgggcgg ttttgaatgt agcaaaatcc tggaggacat caacagctgc
     3721 ctgtggcgct ataaaacaga agaagggcta tatctttgca ccataatttg acctaggctg
     3781 ctgccaccgc tgagcaataa ctagcataac cccttggggc ctctaaacgg gtcttgaggg
     3841 gttttttgct gaaacctcag gcatttgaga agcacacggt cacactgctt ccggtagtca
     3901 ataaaccggt aaaccagcaa tagacataag cggctattta acgaccctgc cctgaaccga
     3961 cgaccgggtc gaatttgctt tcgaatttct gccattcatc cgcttattat cacttattca
     4021 ggcgtagcaa ccaggcgttt aagggcacca ataactgcct taaaaaaatt acgccccgcc
     4081 ctgccactca tcgcagtact gttgtaattc attaagcatt ctgccgacat ggaagccatc
     4141 acaaacggca tgatgaacct gaatcgccag cggcatcagc accttgtcgc cttgcgtata
     4201 atatttgccc atagtgaaaa cgggggcgaa gaagttgtcc atattggcca cgtttaaatc
     4261 aaaactggtg aaactcaccc agggattggc tgacacgaaa aacatattct caataaaccc
     4321 tttagggaaa taggccaggt tttcaccgta acacgccaca tcttgcgaat atatgtgtag
     4381 aaactgccgg aaatcgtcgt ggtattcact ccagagcgat gaaaacgttt cagtttgctc
     4441 atggaaaacg gtgtaacaag ggtgaacact atcccatatc accagctcac cgtctttcat
     4501 tgccatacgg aactccggat gagcattcat caggcgggca agaatgtgaa taaaggccgg
     4561 ataaaacttg tgcttatttt tctttacggt ctttaaaaag gccgtaatat ccagctgaac
     4621 ggtctggtta taggtacatt gagcaactga ctgaaatgcc tcaaaatgtt ctttacgatg
     4681 ccattgggat atatcaacgg tggtatatcc agtgattttt ttctccattt tagcttcctt
     4741 agctcctgaa aatctcgata actcaaaaaa tacgcccggt agtgatctta tttcattatg
     4801 gtgaaagttg gaacctctta cgtgccgatc aacgactcat tttcgccaaa agttggccca
     4861 gggcttcccg gtatcaacag ggacaccagg atttatttat tctgcgaagt gatcttccgt
     4921 cacaggtatt tattcggcgc aaagtgcgtc gggtgatgct gccaacttac tgatttagtg
     4981 tatgatggtg tttttgaggt gctccagtgg cttctgtttc tatcagctgt ccctcctgtt
     5041 cagctactga cggggtggtg cgtaacggca aaagcaccgc cggacatcag cgctagcgga
     5101 gtgtatactg gcttactatg ttggcactga tgagggtgtc agtgaagtgc ttcatgtggc
     5161 aggagaaaaa aggctgcacc ggtgcgtcag cagaatatgt gatacaggat atattccgct
     5221 tcctcgctca ctgactcgct acgctcggtc gttcgactgc ggcgagcgga aatggcttac
     5281 gaacggggcg gagatttcct ggaagatgcc aggaagatac ttaacaggga agtgagaggg
     5341 ccgcggcaaa gccgtttttc cataggctcc gcccccctga caagcatcac gaaatctgac
     5401 gctcaaatca gtggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     5461 gcggctccct cgtgcgctct cctgttcctg cctttcggtt taccggtgtc attccgctgt
     5521 tatggccgcg tttgtctcat tccacgcctg acactcagtt ccgggtaggc agttcgctcc
     5581 aagctggact gtatgcacga accccccgtt cagtccgacc gctgcgcctt atccggtaac
     5641 tatcgtcttg agtccaaccc ggaaagacat gcaaaagcac cactggcagc agccactggt
     5701 aattgattta gaggagttag tcttgaagtc atgcgccggt taaggctaaa ctgaaaggac
     5761 aagttttggt gactgcgctc ctccaagcca gttacctcgg ttcaaagagt tggtagctca
     5821 gagaaccttc gaaaaaccgc cctgcaaggc ggttttttcg ttttcagagc aagagattac
     5881 gcgcagacca aaacgatctc aagaagatca tcttattaat cagataaaat atttctagat
     5941 ttcagtgcaa tttatctctt caaatgtagc acctgaagtc agccccatac gatataagtt
     6001 gtaattctca tgttagtcat gccccgcgcc caccggaagg agctgactgg gttgaaggct
     6061 ctcaagggca tcggtcgaga tcccggtgcc taatgagtga gctaacttac attaattgcg
     6121 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     6181 ggccaacgcg cggggagagg cggtttgcgt attgggcgcc agggtggttt ttcttttcac
     6241 cagtgacacg ggcaacagct gattgccctt caccgcctgg ccctgagaga gttgcagcaa
     6301 gcggtccacg ctggtttgcc ccagcaggcg aaaatcctgt ttgatggtgg ttaacggcgg
     6361 gatataacat gagctgtcct cggtatcgtc gtatcccact accgagatgt ccgcaccaac
     6421 gcgcagcccg gactcggtaa tggcgcgcat tgcgcccagc gccatctgat cgttggcaac
     6481 cagcatcgca gtgggaacga tgccctcatt cagcatttgc atggtttgtt gaaaaccgga
     6541 catggcactc cagtcgcctt cccgttccgc tatcggctga atttgattgc gagtgagata
     6601 tttatgccag ccagccagac gcagacgcgc cgagacagaa cttaatgggc ccgctaacag
     6661 cgcgatttgc tggtgaccca atgcgaccag atgctccacg cccagtcgcg taccgtcctc
     6721 atgggagaaa ataatactgt tgatgggtgt ctggtcagag acatcaagaa ataacgccgg
     6781 aacattagtg caggcagctt ccacagcaat ggcatcctgg tcatccagcg gatagttaat
     6841 gatcagccca ctgacgcgtt gcgcgagaag attgtgcacc gccgctttac aggcttcgac
     6901 gccgcttcgt tctaccatcg acaccaccac gctggcaccc agttgatcgg cgcgagattt
     6961 aatcgccgcg acaatttgcg acggcgcgtg cagggccaga ctggaggtgg caacgccaat
     7021 cagcaacgac tgtttgcccg ccagttgttg tgccacgcgg ttgggaatgt aattcagctc
     7081 cgccatcgcc gcttccactt tttcccgcgt tttcgcagaa acgtggctgg cctggttcac
     7141 cacgcgggaa acggtctgat aagagacacc ggcatactct gcgacatcgt ataacgttac
     7201 tggtttcaca ttcaccaccc tgaattgact ctcttccggg cgctatcatg ccataccgcg
     7261 aaaggttttg cgccattcga tggtgtccg
//
