LOCUS       sequence-399434-        3667 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CyCAST donor harboring non-functional cargo..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3667)
  AUTHORS   Faure G, Saito M, Benler S, Peng I, Wolf YI, Strecker J, Altae-Tran 
            H, Neumann E, Li D, Makarova KS, Macrae RK, Koonin EV, Zhang F
  TITLE     Modularity and diversity of target selectors in Tn7 transposons.
  JOURNAL   Mol Cell. 2023 Jun 15;83(12):2122-2136.e10. doi: 
            10.1016/j.molcel.2023.05.013. Epub 2023 Jun 1.
  PUBMED    37267947
REFERENCE   2  (bases 1 to 3667)
  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.2023.05.013"; journalName: "Mol Cell"; date: 
            "2023-06-15- 15"; volume: "83"; issue: "12"; pages: "2122-2136.e10"
COMMENT     Sequence Label: pMSIP124: pDonor(CyCAST)_0.5kb-cargo
FEATURES             Location/Qualifiers
     source          1..3667
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     364..386
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     378..395
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     379..395
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1446..1462)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1446..1462)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1459..1481)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1470..1486
                     /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)."
     promoter        complement(join(1494..1500,1501..1518,1519..1524))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1539..1560
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1677..1694)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1848..2436)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1928..1947)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2607..3398,3399..3467))
                     /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     3230..3249
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3468..3572)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3640..3658
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagactgtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
      241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
      301 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
      361 tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt gacgcgtatt gggattaaaa
      421 acgttcaacc ccataaatca tgcatgttgt gactgcaacc ataagttgtc tatttcggac
      481 actttatgct tacaattttg gctggacagc ttatgcttac aattttggct ggacagatag
      541 agcgaaagca ataggcaaga cgattatatt tctttgccta taaagtaggc ctattgatgg
      601 atgaccctcc aaccgggttc gtgagtggct tccctgccgc ggccgccgcg ggcgcgggcc
      661 gctcgctcta cggtgggccc gagctcgtgt tccccgaggc catgaaccac cccgcgctga
      721 ccgtgcatcc ggcgcaccag ctgggcgcct ccccgctgca gcccccgcac tccttcttcg
      781 gcgcccagca ccgggaccct ctccatttct acccctgggt cctgcggaac cgcttcttcg
      841 gccaccgctt ccagggtgag tgtccacgct gtgcccgccg aggcggccgg ccggcgcccg
      901 tgctgcggcg atgcggggga ggctcggggg cgcgcggggc tgtttagaag ttactgccgg
      961 gaaggctgca ggtccgcgga ggtagattcc caggcaggga agagctgtgc ggcatccacc
     1021 cgcgccttcg ccgcgtaggt ctccctccca ggaaagcagg tggagatatt ggtgcccttc
     1081 tagagtcaga agtcattcct tcggagcttt tccaggatta aaagcatact tgctcttggc
     1141 aaataaccct aaggcaacac aacggctgct ccaagttcta acttttggac aacttatgct
     1201 ttcgattgtt aacatttggg caacttatgc tttcagtttt ttattttgca aagtgaatat
     1261 tgacgatgac tcaagttgtc cacacccatg agtggacaac ttatgctttc aaaatcggac
     1321 aacttatggt tgcagtcaca catgttgtga ctgggcaagt gagatgatag tcacctgatt
     1381 tgcttctccc tcactgcttc cttagcctaa aatcccaatg gcgcgccgag cttggcgtaa
     1441 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
     1501 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
     1561 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     1621 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctgttc cgcttcctcg
     1681 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     1741 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     1801 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     1861 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     1921 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     1981 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     2041 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     2101 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     2161 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     2221 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     2281 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     2341 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     2401 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     2461 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     2521 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     2581 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     2641 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     2701 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga accacgctca
     2761 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     2821 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     2881 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     2941 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     3001 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     3061 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     3121 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     3181 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     3241 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     3301 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     3361 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     3421 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     3481 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     3541 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     3601 gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc
     3661 ttttgtc
//
