LOCUS       sequence-354377-        3203 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of RoCAST left end, cam, RoCAST right end from Rippkaea 
            orientalis.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3203)
  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 3203)
  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: pRoDonor
FEATURES             Location/Qualifiers
     source          1..3203
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     561..580
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(799..903)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1002..1024
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1016..1033
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1017..1033
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             1336..1995
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1402..1421)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     primer_bind     complement(2124..2140)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2124..2140)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2137..2159)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     rep_origin      complement(2312..2900)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2392..2411)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    complement(3017..3041)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(join(3141..3203,1..729,730..798))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
ORIGIN
        1 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
       61 tgctgcaatg ataccgcgag agccacgctc accggctcca gatttatcag caataaacca
      121 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
      181 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
      241 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
      301 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
      361 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
      421 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
      481 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
      541 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
      601 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
      661 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
      721 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
      781 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
      841 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
      901 gcgcacattt ccccgaaaag tgccagatac ctgaaacaaa acccatcgta cggccaagga
      961 agtctccaat aactgtgatc caccacaagc gccagggttt tcccagtcac gacgttgtaa
     1021 aacgacggcc agtcatgcat aatccgcacg catctggaat aaggaagtgc cattccgcct
     1081 gacctaacat tagcttcgcg gcaaaaacaa actttaaata atagaaagta ttatttaaag
     1141 tttgtttttg ccgtatttta aagtttgttt ttgccactcg attctctgaa accctagtaa
     1201 aatcgttgta ctaaaattct agcatgaccg ctttttcaag tttgagtttg ccgaaatcgt
     1261 tattttcagg tttgacattg ccgaaatagt tgggacttgc tatgatggtt ttatcgtaaa
     1321 tttgtattaa tttgcatgga gaaaaaaatc actggatata ccaccgttga tatatcccaa
     1381 tggcatcgta aagaacattt tgaggcattt cagtcagttg ctcaatgtac ctataaccag
     1441 accgttcagc tggatattac ggccttttta aagaccgtaa agaaaaataa gcacaagttt
     1501 tatccggcct ttattcacat tcttgcccgc ctgatgaatg ctcatccgga gttccgtatg
     1561 gcaatgaaag acggtgagct ggtgatatgg gatagtgttc acccttgtta caccgttttc
     1621 catgagcaaa ctgaaacgtt ttcatcgctc tggagtgaat accacgacga tttccggcag
     1681 tttctacaca tatattcgca agatgtggcg tgttacggtg aaaacctggc ctatttccct
     1741 aaagggttta ttgagaatat gtttttcgtc tcagccaatc cctgggtgag tttcaccagt
     1801 tttgatttaa acgtggccaa tatggacaac ttcttcgccc ccgttttcac tatgggcaaa
     1861 tattatacgc aaggcgacaa ggtgctgatg ccgctggcga ttcaggttca tcatgccgtt
     1921 tgtgatggct tccatgtcgg cagaatgctt aatgaattac aacagtactg cgatgagtgg
     1981 cagggcgggg cgtaattttt ttaaggcagt tattggtgcc ctaagactat ttgacttaaa
     2041 atttttcagg catagctttg ctgtcagatt ttcaggcata accttgccga tgcggswmwr
     2101 msawrstkka wryawgwgtc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc
     2161 agagggcaca atcctattcc gcgctatccg acaatctcca agacattagg tggagttcag
     2221 ttcggcgtat ggcatatgtc gctggaaaga acatgtgagc aaaaggccag caaaaggcca
     2281 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     2341 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     2401 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     2461 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     2521 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     2581 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     2641 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     2701 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     2761 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     2821 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     2881 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctctat
     2941 tcaacaaagc cgccgtcccg tcaagtcagc gtaaatgggt agggggcttc aaatcgtcct
     3001 cgtgatacca attcggagcc tgcttttttg tacaaacttg ttgataatgg caattcaagg
     3061 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     3121 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     3181 tgtctatttc gttcatccat agt
//
