LOCUS       sequence-355121-        8626 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of X. albilineans type I-F1 cas2-3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8626)
  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 8626)
  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: pXalb_IF1_Cas2-3
FEATURES             Location/Qualifiers
     source          1..8626
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             462..653
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(647..669)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    755..894
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     810..829
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(909..926)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1080..1668)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1160..1179)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(1779..1798)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1790..2605
                     /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             1790..2605
                     /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     complement(1863..1882)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      complement(2697..3152)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2834..2855)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3046..3065
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     terminator      complement(3189..3236)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     3232..3250
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(3303..3320)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             6730..6752
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    6767..6791
                     /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)."
     primer_bind     complement(6791..6810)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6792..6810)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6894..6913)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        7119..7196
                     /gene="lacI"
                     /label=lacI promoter
     CDS             7197..8279
                     /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     complement(7216..7235)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 tgcgcatgat cgtgctcctg tcgttgagga cccggctagg ctggcggggt tgccttactg
       61 gttagcagaa tgaatcaccg atacgcgagc gaacgtgaag cgactgctgc tgcaaaacgt
      121 ctgcgacctg agcaacaaca tgaatggtct tcggtttccg tgtttcgtaa agtctggaaa
      181 cgcggaagtc agcgccctgc accattatgt tccggatctg catcgcagga tgctgctggc
      241 taccctgtgg aacacctaca tctgtattaa cgaagcgctg gcattgaccc tgagtgattt
      301 ttctctggtc ccgccgcatc cataccgcca gttgtttacc ctcacaacgt tccagtaacc
      361 gggcatgttc atcatcagta acccgtatcg tgagcatcct ctctcgtttc atcggtatca
      421 ttacccccat gaacagaaat cccccttaca cggaggcatc agtgaccaaa caggaaaaaa
      481 ccgcccttaa catggcccgc tttatcagaa gccagacatt aacgcttctg gagaaactca
      541 acgagctgga cgcggatgaa caggcagaca tctgtgaatc gcttcacgac cacgctgatg
      601 agctttaccg cagctgcctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc
      661 agctcccgga gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc
      721 agggcgcgtc agcgggtgtt ggcgggtgtc ggggcgcagc catgacccag tcacgtagcg
      781 atagcggagt gtatactggc ttaactatgc ggcatcagag cagattgtac tgagagtgca
      841 ccattgcggt gtgaaatacc gcacagatgc gtaaggagaa aataccgcat caggcgctct
      901 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
      961 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     1021 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     1081 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     1141 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     1201 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     1261 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     1321 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     1381 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     1441 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     1501 aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc
     1561 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     1621 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     1681 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     1741 atgaacaata aaactgtctg cttacataaa cagtaataca aggggtgtta tgagccatat
     1801 tcaacgggaa acgtcttgct ctaggccgcg attaaattcc aacatggatg ctgatttata
     1861 tgggtataaa tgggctcgcg ataatgtcgg gcaatcaggt gcgacaatct atcgattgta
     1921 tgggaagccc gatgcgccag agttgtttct gaaacatggc aaaggtagcg ttgccaatga
     1981 tgttacagat gagatggtca gactaaactg gctgacggaa tttatgcctc ttccgaccat
     2041 caagcatttt atccgtactc ctgatgatgc atggttactc accactgcga tccccgggaa
     2101 aacagcattc caggtattag aagaatatcc tgattcaggt gaaaatattg ttgatgcgct
     2161 ggcagtgttc ctgcgccggt tgcattcgat tcctgtttgt aattgtcctt ttaacagcga
     2221 tcgcgtattt cgtctcgctc aggcgcaatc acgaatgaat aacggtttgg ttgatgcgag
     2281 tgattttgat gacgagcgta atggctggcc tgttgaacaa gtctggaaag aaatgcataa
     2341 acttttgcca ttctcaccgg attcagtcgt cactcatggt gatttctcac ttgataacct
     2401 tatttttgac gaggggaaat taataggttg tattgatgtt ggacgagtcg gaatcgcaga
     2461 ccgataccag gatcttgcca tcctatggaa ctgcctcggt gagttttctc cttcattaca
     2521 gaaacggctt tttcaaaaat atggtattga taatcctgat atgaataaat tgcagtttca
     2581 tttgatgctc gatgagtttt tctaagaatt aattcatgag cggatacata tttgaatgta
     2641 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctaaat
     2701 tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt
     2761 taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg
     2821 gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt
     2881 caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc
     2941 aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg
     3001 atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa
     3061 aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc
     3121 cgccgcgctt aatgcgccgc tacagggcgc gtcccattcg ccaatccgga tatagttcct
     3181 cctttcagca aaaaacccct caagacccgt ttagaggccc caaggggtta tgctagttat
     3241 tgctcagcgg tggcagcagc caactcagct tcctttcggg ctttgttagc agccggatct
     3301 cagtggtggt ggtggtggtg ctcttcccac gacttgaagc caagtaaagg gtgatacccc
     3361 cacttcttcc cgcccttgcc ctcaggcacg ttcaccgtcg ccagccgctt ggcacagtcc
     3421 tgcaacgaca ggtcgagctg ctcggcctgc tcggtgagca gcgtcatcag gtcgtattcg
     3481 ccccagggcg cgatgccggg accaggttcg agcgccagcg gcgcgcacgc actctgttcc
     3541 acgaccgggt agagtttttt gtcgccgcga tggtcatcgt cgtcttcgat gcgatgcagc
     3601 accacgcgct cctcgtgctc atcgggtaga aacaccaaag tggccgttgg cctcgtgtcg
     3661 tgacggaacg gctgttgttg cggcagtacg ccggtcagca tggcgtgcgg gtattgccat
     3721 gcgcatgccg catccaagga gaacggcggg gcgctgcgcc gggatggcgg agcggcctcg
     3781 cgcgatttgg gcagcatggc ctcgctcagc cgtgcatgtt ccagatcgca cagccgctct
     3841 tgcggatgcc agggtggctc cggtggttgg atcctgggca gggaggtgag cacgcggtac
     3901 tcgtcctcgc gcaggagttt gccgaggcgg tgggtctgta ggcggaaacg tccagcacgt
     3961 gtgccgtcgg cgtcgttctg ttcgaatccc ggtcgcacat aggccggggc gtcttggtcg
     4021 aggttgcgga aatgcagcag gttggtgtcg aagatcagga tgttcgcggc ggtgccaggc
     4081 gtgcgccgat ggcgttgcac gcgaccaccc aactggatga gcgagcgcag cgaagagggt
     4141 tcggcgatgg cccaatccgc atcccagtcg cgcccgactt cgcagactgg cgaggccagg
     4201 accacgaaca catgcgcctg ttccggatgt gcatcgagcg cggcgcggac gctgggcaag
     4261 cgatacaccg cgtcggcttg gcggcggtcg aatgctgcat cgagcaggtg ttcggtcgct
     4321 gagcgcacca gcagcggaaa gcacgcgtgg tagacgcaca ggtggatgcg gatgccgggt
     4381 gctgcgccga gtcggtacag ttcctgtgcc acctcgatca gcggtcggat attggccatt
     4441 cgcactagac cgaagctgac gtgcttgccg ctgaccgggt cggtttccgc gtgtgcgctg
     4501 tgcaggcgct ggatcgcgtc gcggacatgt ggcgcgaact cggcatggat ctgctccggt
     4561 tgccgcgcct ggatctgtaa ttgaaccaat tcgccgcgtc gcagcggctc ggtcgtgcgc
     4621 aatttggccg cacgttttgt cacgaattgc gcatgctggg cagcgaatgc atcggcatcg
     4681 acgcaatgcg cggtatatgc gttgaactcg tcggtccaca gacaaggcaa ctcgatgcgt
     4741 gcatcggcgc gcccgccatc gacaccgcga ttgcgccgat agtgggcgcg tccggcgcga
     4801 taggcggcga acatgccttc gaccaaggcc ggtgccaacg ttgccgagga caacagcaca
     4861 cgcgtcccga gcaggcccgc ccagtacacc agccgcgtca gggtgggtag gtcgtcgatg
     4921 tcgtagtcgt ccagttcgtc aaggatcagg tcggcgctca tcagccgcag catcggcgcg
     4981 atctggcggc cggcacgcag cgattcggtg gccggtacca tgtggtcgac ggtgcagacc
     5041 agcatcggcg ccgacaacag tttgcgcatc tgcgggtcgt gcagtgcgcg cgacaacaat
     5101 ggatgttcgg tttcggcacc gtcgtagagg acgtggctat cttcctcgat cagcgcttgt
     5161 atcgaggccg agccgcgtgt gtcggcttcg cgttcgtggt gatcgaacag cgggcggctg
     5221 gacgaaccgc cgacgagcat ggcgagcgtg tcgtcgtcga ggtgcagatc gcgccgatag
     5281 ctgtcgccgg tttgtcgggt gagcgtgcgc aggcccagcg cataagtcag acgcaggccc
     5341 agttgcgggt cggccagtgc gtagaggatg cgcgcgttgc ccagggtctt gccgcagccg
     5401 gtggaagcca tgttgacgat gaaggcaccg tgctcgcgtg cggactcgcg cagcgccgtg
     5461 gccgtgtcgg cggccttgtt ctgccaggcg aagtgggcgt cggcgctgcg tttgcgcagg
     5521 ccgcgatgtt gcgccagtcg cggcagagac cgctcgaagc cgggcagcgc atatgcgacc
     5581 aatcccgcgg tatgcgcaac gccgagtaga tgttcgtcca gtgactgatt gcggacgagc
     5641 ttgccggcac ggttgaaggt ggtgttggcg gcaagcgtgg cgtctttttt cagatcctgt
     5701 tgttcgttgc ggtccataac cagacgcgaa tagctgtggt cagccaacat caggctgagc
     5761 cgcgccagat gcatgacgta gggattgtcg agccagcctt cgggtctgcg cacgcgcagt
     5821 gcgtccaagc gtttagccaa ccgcgccgcc tgtgcgcgcc atgctggcgt ggcgaccggc
     5881 aatgggccgg cgagcgtcca gtacgcggcg atcctggcag gttccgcttg cacgcggatc
     5941 tcgttccaat cgtgctcgac cagagacagc ggcagttcca tccaggtcga gatggtgtgc
     6001 tgcacgcgtc gacccagcca gtcctggcga ccgtctgtct tgaagggcgc cagcggcagg
     6061 cggtgatggg tcaggatcaa ccaggcgacg gccgcggcca ggggcggcag ggcgcggaac
     6121 gggtatccgg gctcggcatc cagaccgtcg cggcggtagc gtccttcggc ggtccagcat
     6181 ggcgtatgat cgtgatcggg atcggccagc cgttgcaacc agccggcatc gtcatccttg
     6241 ccgacgaagg cctggaacag gcgtaaagac acccattcgt ggcggtagag gttgcgttct
     6301 ttcaattcgc cgcgcaggcg tgcctggaac gccaggctgg ctttgccgag gtcgtgcagc
     6361 aacgcggcca gttgcgccag caggcgaatg tcttcggcgc agtgccagtc gttttcgtct
     6421 tcgccgcgca aaacatcgcg cgccgtgctg ttggtgggca cggccccctg tgcgttgaat
     6481 cgactggcgt cgccgacgat ccacaacaac tcgctgtggt ccaggccgcg aatccaatga
     6541 caggccaccg cggtgttctt gcgtgcggtg cggcgcagca gtttgcgcaa ggtgtccagg
     6601 ccggcgcggg tgatcggcgt ttgccaggtg cgatcgccgc gccgctcggc gaactggtcc
     6661 aggatgcggc ggctttcttt cagggcacgc ttgtcgcatt gggaaaccag caagatattc
     6721 atcggtatat ctccttctta aagttaaaca aaattatttc tagaggggaa ttgttatccg
     6781 ctcacaattc ccctatagtg agtcgtatta atttcgcggg atcgagatct cgatcctcta
     6841 cgccggacgc atcgtggccg gcatcaccgg cgccacaggt gcggttgctg gcgcctatat
     6901 cgccgacatc accgatgggg aagatcgggc tcgccacttc gggctcatga gcgcttgttt
     6961 cggcgtgggt atggtggcag gccccgtggc cgggggactg ttgggcgcca tctccttgca
     7021 tgcaccattc cttgcggcgg cggtgctcaa cggcctcaac ctactactgg gctgcttcct
     7081 aatgcaggag tcgcataagg gagagcgtcg agatcccgga caccatcgaa tggcgcaaaa
     7141 cctttcgcgg tatggcatga tagcgcccgg aagagagtca attcagggtg gtgaatgtga
     7201 aaccagtaac gttatacgat gtcgcagagt atgccggtgt ctcttatcag accgtttccc
     7261 gcgtggtgaa ccaggccagc cacgtttctg cgaaaacgcg ggaaaaagtg gaagcggcga
     7321 tggcggagct gaattacatt cccaaccgcg tggcacaaca actggcgggc aaacagtcgt
     7381 tgctgattgg cgttgccacc tccagtctgg ccctgcacgc gccgtcgcaa attgtcgcgg
     7441 cgattaaatc tcgcgccgat caactgggtg ccagcgtggt ggtgtcgatg gtagaacgaa
     7501 gcggcgtcga agcctgtaaa gcggcggtgc acaatcttct cgcgcaacgc gtcagtgggc
     7561 tgatcattaa ctatccgctg gatgaccagg atgccattgc tgtggaagct gcctgcacta
     7621 atgttccggc gttatttctt gatgtctctg accagacacc catcaacagt attattttct
     7681 cccatgaaga cggtacgcga ctgggcgtgg agcatctggt cgcattgggt caccagcaaa
     7741 tcgcgctgtt agcgggccca ttaagttctg tctcggcgcg tctgcgtctg gctggctggc
     7801 ataaatatct cactcgcaat caaattcagc cgatagcgga acgggaaggc gactggagtg
     7861 ccatgtccgg ttttcaacaa accatgcaaa tgctgaatga gggcatcgtt cccactgcga
     7921 tgctggttgc caacgatcag atggcgctgg gcgcaatgcg cgccattacc gagtccgggc
     7981 tgcgcgttgg tgcggatatc tcggtagtgg gatacgacga taccgaagac agctcatgtt
     8041 atatcccgcc gttaaccacc atcaaacagg attttcgcct gctggggcaa accagcgtgg
     8101 accgcttgct gcaactctct cagggccagg cggtgaaggg caatcagctg ttgcccgtct
     8161 cactggtgaa aagaaaaacc accctggcgc ccaatacgca aaccgcctct ccccgcgcgt
     8221 tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag
     8281 cgcaacgcaa ttaatgtaag ttagctcact cattaggcac cgggatctcg accgatgccc
     8341 ttgagagcct tcaacccagt cagctccttc cggtgggcgc ggggcatgac tatcgtcgcc
     8401 gcacttatga ctgtcttctt tatcatgcaa ctcgtaggac aggtgccggc agcgctctgg
     8461 gtcattttcg gcgaggaccg ctttcgctgg agcgcgacga tgatcggcct gtcgcttgcg
     8521 gtattcggaa tcttgcacgc cctcgctcaa gccttcgtca ctggtcccgc caccaaacgt
     8581 ttcggcgaga agcaggccat tatcgccggc atggcggccc cacggg
//
