LOCUS       sequence-333888-        3501 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  PmcCAST donor harboring non-functional cargo..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3501)
  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 3501)
  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.cell.2021.03.006"; journalName: "Cell"; date: 
            "2021-04-29- 29"; volume: "184"; issue: "9"; pages: "2441-2453.e18"
COMMENT     Sequence Label: pMS29: pDonor(PmcCAST)
FEATURES             Location/Qualifiers
     source          1..3501
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     179..198
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     440..457
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    574..595
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(610..615,616..633,634..640)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    648..664
                     /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     653..675
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     672..688
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     672..688
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1565..1582)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1565..1581)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1574..1596)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(1790..1809)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1909..1931
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1969..1987)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2055..2159
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2160..2228,2229..3020)
                     /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(2378..2397)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
       61 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
      121 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
      181 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
      241 ttttgtgatt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
      301 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
      361 atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg
      421 cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg
      481 caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc
      541 cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg agttagctca ctcattaggc
      601 accccaggct ttacacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata
      661 acaatttcac acaggaaaca gctatgacca tgattacgcc aagctcggcg cgccattggg
      721 attgtgactt tacccataac tttgccgcaa cggcaagctt atgcttccaa taaagggtgg
      781 caaggttatg gttccagtta tataagctgt tctagtttta catataacag cttacaaacc
      841 aaactcacgt cgtggactct agaagggcac caatatctcc acctgctttc ctgggaggga
      901 gacctacgcg gcgaaggcgc gggtggatgc cgcacagctc ttccctgcct gggaatctac
      961 ctccgcggac ctgcagcctt cccggcagta acttctaaac agccccgcgc gcccccgagc
     1021 ctcccccgca tcgccgcagc acgggcgccg gccggccgcc tcggcgggca cagcgtggac
     1081 actcaccctg gaagcggtgg ccgaagaagc ggttccgcag gacccagggg tagaaatgga
     1141 gagggtcccg gtgctgggcg ccgaagaagg agtgcggggg ctgcagcggg gaggcgccca
     1201 gctggtgcgc cggatgcacg gtcagcgcgg ggtggttcat ggcctcgggg aacacgagct
     1261 cgggcccacc gtagagcgag cggcccgcgc ccgcggcggc cgcggcaggg aagccactca
     1321 cgaacccggc ttctgctcca ttctaaaaaa attcggcgtt ttcctgcttt ctatgcttga
     1381 ttggactcac ggcaagctta tccttccaat ttggactagc ggcaaagtta tggttccaaa
     1441 ctgtcaattt catccaaaat tagctgacgc aagaatgagg gttgtagtgt tgttgacggc
     1501 aaagttatgg ttccagtaac ggcaaagtta tgggtaaagt cacaatccca atacgcgtca
     1561 attcactggc cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt acccaactta
     1621 atcgccttgc agcacatccc cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg
     1681 atcgcccttc ccaacagttg cgcagcctga atggcgaatg gcgcctgatg cggtattttc
     1741 tccttacgca tctgtgcggt atttcacacc gcatatggtg cactctcagt acaatctgct
     1801 ctgatgccgc atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac
     1861 gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca
     1921 tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac
     1981 gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt
     2041 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     2101 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
     2161 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     2221 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac
     2281 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
     2341 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
     2401 gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
     2461 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
     2521 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
     2581 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
     2641 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
     2701 ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
     2761 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
     2821 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
     2881 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
     2941 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
     3001 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt
     3061 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     3121 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     3181 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     3241 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     3301 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
     3361 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     3421 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     3481 taccggataa ggcgcagcgg t
//
