LOCUS       sequence-330860-        3572 bp    DNA     circular SYN 14-APR-2025
DEFINITION  AvCAST donor harboring non-functional cargo..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3572)
  AUTHORS   Saito M, Ladha A, Strecker J, Faure G, Neumann E, Altae-Tran H, 
            Macrae RK, Zhang F
  TITLE     Dual modes of CRISPR-associated transposon homing.
  JOURNAL   Cell. 2021 Apr 29;184(9):2441-2453.e18. doi: 
            10.1016/j.cell.2021.03.006. Epub 2021 Mar 25.
  PUBMED    33770501
REFERENCE   2  (bases 1 to 3572)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1016/j.cell.2021.03.006"; journalName: "Cell"; date: 
            "2021-04-29- 29"; volume: "184"; issue: "9"; pages: "2441-2453.e18"
COMMENT     Sequence Label: pMS12: pDonor(AvCAST)
FEATURES             Location/Qualifiers
     source          1..3572
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     133..152
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     386..403
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    520..541
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(556..561,562..579,580..586)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    594..610
                     /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     599..621
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     618..634
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     618..634
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1590..1607)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1590..1606)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1599..1621)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(1815..1834)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1934..1956
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1994..2012)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2080..2184
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2185..2253,2254..3045)
                     /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"
     primer_bind     complement(2403..2422)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(3216..3572,1..232)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
       61 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
      121 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
      181 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
      241 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
      301 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc
      361 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc
      421 aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag
      481 gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agctcactca
      541 ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag
      601 cggataacaa tttcacacag gaaacagcta tgaccatgat tacgccaagc ttgtggagga
      661 ataattcggt tcaacaatat ataattcggt tcaacaatta ataaaacgat tgaacaataa
      721 cctctaaagg agtacgattg taccaagtaa aagtgcgatc gcttccagag gttattttta
      781 tggttagggg tagaagggaa tggcgggttc gtgagtggct tccctgccgc ggccgccgcg
      841 ggcgcgggcc gctcgctcta cggtgggccc gagctcgtgt tccccgaggc catgaaccac
      901 cccgcgctga ccgtgcatcc ggcgcaccag ctgggcgcct ccccgctgca gcccccgcac
      961 tccttcttcg gcgcccagca ccgggaccct ctccatttct acccctgggt cctgcggaac
     1021 cgcttcttcg gccaccgctt ccagggtgag tgtccacgct gtgcccgccg aggcggccgg
     1081 ccggcgcccg tgctgcggcg atgcggggga ggctcggggg cgcgcggggc tgtttagaag
     1141 ttactgccgg gaaggctgca ggtccgcgga ggtagattcc caggcaggga agagctgtgc
     1201 ggcatccacc cgcgccttcg ccgcgtaggt ctccctccca ggaaagcagg tggagatatt
     1261 ggtgcccttc tagagtcccg ggatatttac gttttgtaat cacgaataga ttcttgattg
     1321 caagatgaat tgcgatcgca tctttgtcgc acagcagaac tttctcaaat aattttgaac
     1381 ttatggaatt actcaaaacc tttatcagta atggattcaa acaattgttt aaccgtttta
     1441 tttgagattt ttactaaaaa tcaagtttac catagaggtt aagctgaaat tgttcaatcg
     1501 ttttatttta tcgacaacgg cagattaaat attcgaactg taataattgt ttaaccgaat
     1561 tctttgtcta cagtaccgag ctcgaattca ctggccgtcg ttttacaacg tcgtgactgg
     1621 gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt cgccagctgg
     1681 cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc
     1741 gaatggcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata
     1801 tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagcc ccgacacccg
     1861 ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa
     1921 gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc
     1981 gcgagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg
     2041 gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     2101 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     2161 caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc
     2221 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa
     2281 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt
     2341 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt
     2401 ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc
     2461 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg
     2521 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg
     2581 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac
     2641 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca
     2701 aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta
     2761 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat
     2821 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa
     2881 tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag
     2941 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat
     3001 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt
     3061 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     3121 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     3181 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     3241 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     3301 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     3361 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     3421 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     3481 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     3541 ggttcgtgca cacagcccag cttggagcga ac
//
