LOCUS       sequence-234055-        7091 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses wildtype V-C Cas1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7091)
  AUTHORS   Wright AV, Wang JY, Burstein D, Harrington LB, Paez-Espino D, 
            Kyrpides NC, Iavarone AT, Banfield JF, Doudna JA
  TITLE     A Functional Mini-Integrase in a Two-Protein-type V-C CRISPR System.
  JOURNAL   Mol Cell. 2019 Feb 21;73(4):727-737.e3. doi: 
            10.1016/j.molcel.2018.12.015. Epub 2019 Jan 29.
  PUBMED    30709710
REFERENCE   2  (bases 1 to 7091)
  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"; doi: 
            "10.1016/j.molcel.2018.12.015"; journalName: "Mol Cell"; date: 
            "2019-02-21- 21"; volume: "73"; issue: "4"; pages: "727-737.e3"
COMMENT     Sequence Label: pET-3a-His-MBP-TEV-VC_Cas1_WT
FEATURES             Location/Qualifiers
     source          1..7091
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1018..1041
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     CDS             1099..1119
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     primer_bind     complement(2376..2394)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2390..2437
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(2803..2808,2809..2825,2826..2831))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(2858..2876)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2944..3048
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3049..3117,3118..3909)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(3267..3286)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4080..4668
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4569..4588
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4822..4839
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4854..4993
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4919..4938)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5079..5101
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(5095..5286)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     6744..6763
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     6847..6866
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6847..6865
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             6954..6976
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             6996..7025
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             join(7035..7091,1..1044)
                     /codon_start=1
                     /gene="malE (mutated)"
                     /product="maltose binding protein from E. coli"
                     /label=MBP
                     /note="This version of the gene does not encode a signal 
                     sequence, so MBP will remain in the cytosol."
                     /translation="MKIEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKLE
                     EKFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNGKL
                     IAYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWPLIA
                     ADGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAFNKGE
                     TAMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKELAKEFL
                     ENYLLTDEGLEAVNKDKPLGAVALKSYEEELAKDPRIAATMENAQKGEIMPNIPQMSAF
                     WYAVRTAVINAASGRQTVDEALKDAQT"
ORIGIN
        1 ggtctcgctg aagtcggtaa gaaattcgag aaagataccg gaattaaagt caccgttgag
       61 catccggata aactggaaga gaaattccca caggttgcgg caactggcga tggccctgac
      121 attatcttct gggcacacga ccgctttggt ggctacgctc aatctggcct gttggctgaa
      181 atcaccccgg acaaagcgtt ccaggacaag ctgtatccgt ttacctggga tgccgtacgt
      241 tacaacggca agctgattgc ttacccgatc gctgttgaag cgttatcgct gatttataac
      301 aaagatctgc tgccgaaccc gccaaaaacc tgggaagaga tcccggcgct ggataaagaa
      361 ctgaaagcga aaggtaagag cgcgctgatg ttcaacctgc aagaaccgta cttcacctgg
      421 ccgctgattg ctgctgacgg gggttatgcg ttcaagtatg aaaacggcaa gtacgacatt
      481 aaagacgtgg gcgtggataa cgctggcgcg aaagcgggtc tgaccttcct ggttgacctg
      541 attaaaaaca aacacatgaa tgcagacacc gattactcca tcgcagaagc tgcctttaat
      601 aaaggcgaaa cagcgatgac catcaacggc ccgtgggcat ggtccaacat cgacaccagc
      661 aaagtgaatt atggtgtaac ggtactgccg accttcaagg gtcaaccatc caaaccgttc
      721 gttggcgtgc tgagcgcagg tattaacgcc gccagtccga acaaagagct ggcaaaagag
      781 ttcctcgaaa actatctgct gactgatgaa ggtctggaag cggttaataa agacaaaccg
      841 ctgggtgccg tagcgctgaa gtcttacgag gaagagttgg cgaaagatcc acgtattgcc
      901 gccactatgg aaaacgccca gaaaggtgaa atcatgccga acatcccgca gatgtccgct
      961 ttctggtatg ccgtgcgtac tgcggtgatc aacgccgcca gcggtcgtca gactgtcgat
     1021 gaagccctga aagacgcgca gactaattcg agctcgaaca acaacaacaa taacaataac
     1081 aacaacctcg ggatcgagga aaacctgtac ttccaatcca atgcaatgga tcagggtaat
     1141 cagaccattg aaaaccagac cattaattgt gatcagcacc cggattttct gtggacctgg
     1201 aaaagcaata aacgtggtag ccgtgttagc gtttggctgc cgtattttag ccaggccaaa
     1261 aaaatccctc gtagcaaaaa atggtcagtg gcatataatg gtggcagcat tgaattcgat
     1321 ctgaaagaaa ccgatctgat catgttttat ggtgcaaccg gtgaactgcc gctggaattt
     1381 ctggatgatg caagcaaaaa tggtgtgatg attctgattc atcgtcgtaa tgttctgcag
     1441 ccgtatgttt tttatccgag cgttattggt gatgaagaag atattctgac caagcagatt
     1501 cagtttcgta ccaatgaacg taaacgtctg tatattgcaa agaccctgat caaaaaacgc
     1561 ctggaaaata tgggtagcac cattccgatt agcgcaccgc tgctgcgtca gctgagtgca
     1621 gcaaaaagca ttgatgaagt tcgtgcaatt gaagcaaata ccaccgcacg ttattggaac
     1681 aaatggtatg aaaacctgaa catcgaaacc acgcgtcgta aagatcatcc gattaatagc
     1741 gcactggatg caggtagcaa atttatctat ggtgttattc tgcgttggct ggtgtttcat
     1801 cgttttagcc cgaatcatgg ctttatgcat cagccgacca gctatccgag cctggtttat
     1861 gatctgatgg aaccgtttcg ctatatgatt gaaaatgttt gtagcgcagc atggaaacgt
     1921 ggtgaacgtg aaaatagcaa aattgttgca ctgagcctga gctttctgaa agaggaactg
     1981 gataaaccgt gttatgttcc ggcaacacgt cagtatgttc gcaaaaaaaa cctgctgcat
     2041 ggtgcagttc tggcactgcg tagctatctg attggtgata tgcgtaaact ggtttttccg
     2101 agcgaaggtg ttccgaatgg tggtcgtccg attaaagcaa gctataaact gcctggtagc
     2161 atgtatgatg ttggtcgtaa accgcctgaa atcaaacaga aagatgaaat ctgctttgac
     2221 gaagtgagcc aagaagaaag cgaagagtaa taacattgga agtggataac ggatccgcga
     2281 tcgcggcgcg ccacctggtg gccggccggt accacgcgtg cgcgctgatc cggctgctaa
     2341 caaagcccga aaggaagctg agttggctgc tgccaccgct gagcaataac tagcataacc
     2401 ccttggggcc tctaaacggg tcttgagggg ttttttgctg aaaggaggaa ctatatccgg
     2461 atatccacag gacgggtgtg gtcgccatga tcgcgtagtc gatagtggct ccaagtagcg
     2521 aagcgagcag gactgggcgg cggccaaagc ggtcggacag tgctccgaga acgggtgcgc
     2581 atagaaattg catcaacgca tatagcgcta gcagcacgcc atagtgactg gcgatgctgt
     2641 cggaatggac gatatcccgc aagaggcccg gcagtaccgg cataaccaag cctatgccta
     2701 cagcatccag ggtgacggtg ccgaggatga cgatgagcgc attgttagat ttcatacacg
     2761 gtgcctgact gcgttagcaa tttaactgtg ataaactacc gcattaaagc ttatcgatga
     2821 taagctgtca aacatgagaa ttcttgaaga cgaaagggcc tcgtgatacg cctattttta
     2881 taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat
     2941 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     3001 agacaataac cctgataaat gcttcaataa cattgaaaaa ggaagagtat gagtattcaa
     3061 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
     3121 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
     3181 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
     3241 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tgttgacgcc
     3301 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
     3361 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
     3421 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
     3481 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
     3541 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgcagcaatg
     3601 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
     3661 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
     3721 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt
     3781 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
     3841 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
     3901 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat
     3961 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     4021 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     4081 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     4141 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     4201 agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc
     4261 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     4321 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     4381 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     4441 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     4501 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     4561 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     4621 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     4681 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     4741 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
     4801 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg
     4861 cggtattttc tccttacgca tctgtgcggt atttcacacc gcaatggtgc actctcagta
     4921 caatctgctc tgatgccgca tagttaagcc agtatacact ccgctatcgc tacgtgactg
     4981 ggtcatggct gcgccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct
     5041 gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag
     5101 gttttcaccg tcatcaccga aacgcgcgag gcagctgcgg taaagctcat cagcgtggtc
     5161 gtgaagcgat tcacagatgt ctgcctgttc atccgcgtcc agctcgttga gtttctccag
     5221 aagcgttaat gtctggcttc tgataaagcg ggccatgtta agggcggttt tttcctgttt
     5281 ggtcactgat gcctccgtgt aagggggatt tctgttcatg ggggtaatga taccgatgaa
     5341 acgagagagg atgctcacga tacgggttac tgatgatgaa catgcccggt tactggaacg
     5401 ttgtgagggt aaacaactgg cggtatggat gcggcgggac cagagaaaaa tcactcaggg
     5461 tcaatgccag cgcttcgtta atacagatgt aggtgttcca cagggtagcc agcagcatcc
     5521 tgcgatgcag atccggaaca taatggtgca gggcgctgac ttccgcgttt ccagacttta
     5581 cgaaacacgg aaaccgaaga ccattcatgt tgttgctcag gtcgcagacg ttttgcagca
     5641 gcagtcgctt cacgttcgct cgcgtatcgg tgattcattc tgctaaccag taaggcaacc
     5701 ccgccagcct agccgggtcc tcaacgacag gagcacgatc atgcgcaccc gtggccagga
     5761 cccaacgctg cccgagatgc gccgcgtgcg gctgctggag atggcggacg cgatggatat
     5821 gttctgccaa gggttggttt gcgcattcac agttctccgc aagaattgat tggctccaat
     5881 tcttggagtg gtgaatccgt tagcgaggtg ccgccggctt ccattcaggt cgaggtggcc
     5941 cggctccatg caccgcgacg caacgcgggg aggcagacaa ggtatagggc ggcgcctaca
     6001 atccatgcca acccgttcca tgtgctcgcc gaggcggcat aaatcgccgt gacgatcagc
     6061 ggtccaatga tcgaagttag gctggtaaga gccgcgagcg atccttgaag ctgtccctga
     6121 tggtcgtcat ctacctgcct ggacagcatg gcctgcaacg cgggcatccc gatgccgccg
     6181 gaagcgagaa gaatcataat ggggaaggcc atccagcctc gcgtcgcgaa cgccagcaag
     6241 acgtagccca gcgcgtcggc cgccatgccg gcgataatgg cctgcttctc gccgaaacgt
     6301 ttggtggcgg gaccagtgac gaaggcttga gcgagggcgt gcaagattcc gaataccgca
     6361 agcgacaggc cgatcatcgt cgcgctccag cgaaagcggt cctcgccgaa aatgacccag
     6421 agcgctgccg gcacctgtcc tacgagttgc atgataaaga agacagtcat aagtgcggcg
     6481 acgatagtca tgccccgcgc ccaccggaag gagctgactg ggttgaaggc tctcaagggc
     6541 atcggtcgac gctctccctt atgcgactcc tgcattagga agcagcccag tagtaggttg
     6601 aggccgttga gcaccgccgc cgcaaggaat ggtgcatgca aggagatggc gcccaacagt
     6661 cccccggcca cggggcctgc caccataccc acgccgaaac aagcgctcat gagcccgaag
     6721 tggcgagccc gatcttcccc atcggtgatg tcggcgatat aggcgccagc aaccgcacct
     6781 gtggcgccgg tgatgccggc cacgatgcgt ccggcgtaga ggatcgagat ctcgatcccg
     6841 cgaaattaat acgactcact atagggagac cacaacggtt tccctctagt gccggctccg
     6901 gagagctctt taattaagcg gccgccctgc aggactcgag ttctagaaat aattttgttt
     6961 aactttaaga aggagatata catatgaaat cttctcacca tcaccatcac catcaccatc
     7021 accatggttc ttctatgaaa atcgaagaag gtaaactggt aatctggatt aacggcgata
     7081 aaggctataa c
//
