LOCUS       sequence-355153-        6261 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of A. chroococcum type I-E #3 cas7.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6261)
  AUTHORS   Wimmer F, Mougiakos I, Englert F, Beisel CL
  TITLE     Rapid cell-free characterization of multi-subunit CRISPR effectors 
            and transposons.
  JOURNAL   Mol Cell. 2022 Feb 16. pii: S1097-2765(22)00102-2. doi: 
            10.1016/j.molcel.2022.01.026.
  PUBMED    35216669
REFERENCE   2  (bases 1 to 6261)
  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: "Mol Cell. 
            2022 Feb 16. pii: S1097-2765(22)00102-2. doi: 
            10.1016/j.molcel.2022.01.026."
COMMENT     Sequence Label: pAc3_Cas7
FEATURES             Location/Qualifiers
     source          1..6261
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(775..1857)
                     /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     1819..1838
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(1858..1935)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     2141..2160
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     2244..2263
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2244..2262
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2263..2287
                     /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             3369..3386
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3439..3457)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3453..3500
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      3537..3992
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3624..3643)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3834..3855
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(4084..4899)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(4084..4899)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     4807..4826
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     4891..4910
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      5021..5609
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5510..5529
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5763..5780
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    5795..5934
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(5860..5879)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6020..6042
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(6036..6227)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 ggggtaatga taccgatgaa acgagagagg atgctcacga tacgggttac tgatgatgaa
       61 catgcccggt tactggaacg ttgtgagggt aaacaactgg cggtatggat gcggcgggac
      121 cagagaaaaa tcactcaggg tcaatgccag cgcttcgtta atacagatgt aggtgttcca
      181 cagggtagcc agcagcatcc tgcgatgcag atccggaaca taatggtgca gggcgctgac
      241 ttccgcgttt ccagacttta cgaaacacgg aaaccgaaga ccattcatgt tgttgctcag
      301 gtcgcagacg ttttgcagca gcagtcgctt cacgttcgct cgcgtatcgg tgattcattc
      361 tgctaaccag taaggcaacc ccgccagcct agccgggtcc tcaacgacag gagcacgatc
      421 atgcgcaccc gtggggccgc catgccggcg ataatggcct gcttctcgcc gaaacgtttg
      481 gtggcgggac cagtgacgaa ggcttgagcg agggcgtgca agattccgaa taccgcaagc
      541 gacaggccga tcatcgtcgc gctccagcga aagcggtcct cgccgaaaat gacccagagc
      601 gctgccggca cctgtcctac gagttgcatg ataaagaaga cagtcataag tgcggcgacg
      661 atagtcatgc cccgcgccca ccggaaggag ctgactgggt tgaaggctct caagggcatc
      721 ggtcgagatc ccggtgccta atgagtgagc taacttacat taattgcgtt gcgctcactg
      781 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
      841 gggagaggcg gtttgcgtat tgggcgccag ggtggttttt cttttcacca gtgagacggg
      901 caacagctga ttgcccttca ccgcctggcc ctgagagagt tgcagcaagc ggtccacgct
      961 ggtttgcccc agcaggcgaa aatcctgttt gatggtggtt aacggcggga tataacatga
     1021 gctgtcttcg gtatcgtcgt atcccactac cgagatatcc gcaccaacgc gcagcccgga
     1081 ctcggtaatg gcgcgcattg cgcccagcgc catctgatcg ttggcaacca gcatcgcagt
     1141 gggaacgatg ccctcattca gcatttgcat ggtttgttga aaaccggaca tggcactcca
     1201 gtcgccttcc cgttccgcta tcggctgaat ttgattgcga gtgagatatt tatgccagcc
     1261 agccagacgc agacgcgccg agacagaact taatgggccc gctaacagcg cgatttgctg
     1321 gtgacccaat gcgaccagat gctccacgcc cagtcgcgta ccgtcttcat gggagaaaat
     1381 aatactgttg atgggtgtct ggtcagagac atcaagaaat aacgccggaa cattagtgca
     1441 ggcagcttcc acagcaatgg catcctggtc atccagcgga tagttaatga tcagcccact
     1501 gacgcgttgc gcgagaagat tgtgcaccgc cgctttacag gcttcgacgc cgcttcgttc
     1561 taccatcgac accaccacgc tggcacccag ttgatcggcg cgagatttaa tcgccgcgac
     1621 aatttgcgac ggcgcgtgca gggccagact ggaggtggca acgccaatca gcaacgactg
     1681 tttgcccgcc agttgttgtg ccacgcggtt gggaatgtaa ttcagctccg ccatcgccgc
     1741 ttccactttt tcccgcgttt tcgcagaaac gtggctggcc tggttcacca cgcgggaaac
     1801 ggtctgataa gagacaccgg catactctgc gacatcgtat aacgttactg gtttcacatt
     1861 caccaccctg aattgactct cttccgggcg ctatcatgcc ataccgcgaa aggttttgcg
     1921 ccattcgatg gtgtccggga tctcgacgct ctcccttatg cgactcctgc attaggaagc
     1981 agcccagtag taggttgagg ccgttgagca ccgccgccgc aaggaatggt gcatgcaagg
     2041 agatggcgcc caacagtccc ccggccacgg ggcctgccac catacccacg ccgaaacaag
     2101 cgctcatgag cccgaagtgg cgagcccgat cttccccatc ggtgatgtcg gcgatatagg
     2161 cgccagcaac cgcacctgtg gcgccggtga tgccggccac gatgcgtccg gcgtagagga
     2221 tcgagatctc gatcccgcga aattaatacg actcactata ggggaattgt gagcggataa
     2281 caattccctc tagataactc atatttgaat agctagggag gtttattatg actagatttg
     2341 tgcagcttca tcttctcatc tcttatccac cagccaatct gaatcgtgat gacttgggtc
     2401 agcctaaaac ggcaagactt gggggcgtcg aacgtctgcg tgtgagttct caaagcctca
     2461 aaagagcgtg gcgcacatcc gagctgtttc agcagcagtt ggctggatca attggcactc
     2521 gtactaaacg tctcggtaga gaagctttcg atgcactcat tgctgccggc ataggcgaaa
     2581 agcaggctac agaatgggcg agccagattg ccaaagtgta tggagcagta aaaaaggata
     2641 atccgttaga gattgagcag ttagtacata tagcaccgga agagcgcgca tcactaggat
     2701 cgctggttgc aattctcgcc cgtgaaaaaa gagccccatc ggacgaagag ctggatgcgc
     2761 ttctacacaa gcagtcggct gtagacattg cgatgtttgg tcgtatgctg gctgcaaaga
     2821 cggaatacaa cggggatgcg gcggttcagg ttgcacacgc aattggtgtt catgcaagtg
     2881 ctatagaaga tgattatttt accgcagtcg atgacctgaa tacggatgac ccgggtgcag
     2941 cacacattgg tgaatccggt tttgcggcgg ccgtgtttta tcagtatctt tgcatcgata
     3001 gagatctgtt gaaacgtaac cttggcggtg acgaggaact gactcagaaa gccctgatag
     3061 ctttggctga agcagcgctg aaagtgggcc cgagcggcaa gcaaaactca tttgcttcac
     3121 gagcatacgc ccattatgca ctcgccgaaa agggtcctca gcagccccgg tccttaagcc
     3181 tggccttcat taagcctgtg tctgggggcg actatgcatc agaagccgtc gaagtcttgc
     3241 agcgtttgcg cgacaacatg gataaggtgt atggggcatg cgcggattca cgttatcaat
     3301 ttgatgtttt aaaaggagag ggttctcttg cccaactgtt agattttgtg gcagccgagt
     3361 aactcgagca ccaccaccac caccactgag atccggctgc taacaaagcc cgaaaggaag
     3421 ctgagttggc tgctgccacc gctgagcaat aactagcata accccttggg gcctctaaac
     3481 gggtcttgag gggttttttg ctgaaaggag gaactatatc cggattggcg aatgggacgc
     3541 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     3601 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     3661 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
     3721 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc
     3781 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     3841 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg
     3901 gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     3961 gaattttaac aaaatattaa cgcttacaat ttaggtggca cttttcgggg aaatgtgcgc
     4021 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgaattaa
     4081 ttcttagaaa aactcatcga gcatcaaatg aaactgcaat ttattcatat caggattatc
     4141 aataccatat ttttgaaaaa gccgtttctg taatgaagga gaaaactcac cgaggcagtt
     4201 ccataggatg gcaagatcct ggtatcggtc tgcgattccg actcgtccaa catcaataca
     4261 acctattaat ttcccctcgt caaaaataag gttatcaagt gagaaatcac catgagtgac
     4321 gactgaatcc ggtgagaatg gcaaaagttt atgcatttct ttccagactt gttcaacagg
     4381 ccagccatta cgctcgtcat caaaatcact cgcatcaacc aaaccgttat tcattcgtga
     4441 ttgcgcctga gcgagacgaa atacgcgatc gctgttaaaa ggacaattac aaacaggaat
     4501 cgaatgcaac cggcgcagga acactgccag cgcatcaaca atattttcac ctgaatcagg
     4561 atattcttct aatacctgga atgctgtttt cccggggatc gcagtggtga gtaaccatgc
     4621 atcatcagga gtacggataa aatgcttgat ggtcggaaga ggcataaatt ccgtcagcca
     4681 gtttagtctg accatctcat ctgtaacatc attggcaacg ctacctttgc catgtttcag
     4741 aaacaactct ggcgcatcgg gcttcccata caatcgatag attgtcgcac ctgattgccc
     4801 gacattatcg cgagcccatt tatacccata taaatcagca tccatgttgg aatttaatcg
     4861 cggcctagag caagacgttt cccgttgaat atggctcata acaccccttg tattactgtt
     4921 tatgtaagca gacagtttta ttgttcatga ccaaaatccc ttaacgtgag ttttcgttcc
     4981 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     5041 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     5101 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     5161 atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     5221 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     5281 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     5341 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     5401 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     5461 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     5521 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     5581 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     5641 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
     5701 ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc
     5761 gcagcgagtc agtgagcgag gaagcggaag agcgcctgat gcggtatttt ctccttacgc
     5821 atctgtgcgg tatttcacac cgcaatggtg cactctcagt acaatctgct ctgatgccgc
     5881 atagttaagc cagtatacac tccgctatcg ctacgtgact gggtcatggc tgcgccccga
     5941 cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac
     6001 agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg
     6061 aaacgcgcga ggcagctgcg gtaaagctca tcagcgtggt cgtgaagcga ttcacagatg
     6121 tctgcctgtt catccgcgtc cagctcgttg agtttctcca gaagcgttaa tgtctggctt
     6181 ctgataaagc gggccatgtt aagggcggtt ttttcctgtt tggtcactga tgcctccgtg
     6241 taagggggat ttctgttcat g
//
