LOCUS       sequence-399456-        2753 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Reporter for ZooTsy excision..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2753)
  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 2753)
  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: pSCT375: pDonor(ZooTsy)_excision-reporter
FEATURES             Location/Qualifiers
     source          1..2753
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(103..122)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             114..923
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(181..200)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      1101..1689
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1590..1609
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1843..1860
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2124..2145
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2160..2165,2166..2183,2184..2190)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2198..2214
                     /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     2203..2225
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2222..2238
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2222..2238
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             2234..2557
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCTLSTICS
                     DAA"
     misc_feature    2251..2307
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(2308..2325)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2308..2324)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2317..2339)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(2533..2552)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2703..2725
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 tgcagctctg gcccgtgtct caaaatctct gatgttacat tgcacaagat aaaaatatat
       61 catcatgaac aataaaactg tctgcttaca taaacagtaa tacaaggggt gttatgagcc
      121 atattcaacg ggaaacgtcg aggccgcgat taaattccaa catggatgct gatttatatg
      181 ggtataaatg ggctcgcgat aatgtcgggc aatcaggtgc gacaatctat cgcttgtatg
      241 ggaagcccga tgcgccagag ttgtttctga aacatggcaa aggtagcgtt gccaatgatg
      301 ttacagatga gatggtcaga ctaaactggc tgacggaatt tatgcctctt ccgaccatca
      361 agcattttat ccgtactcct gatgatgcat ggttactcac cactgcgatc cccggaaaaa
      421 cagcattcca ggtattagaa gaatatcctg attcaggtga aaatattgtt gatgcgctgg
      481 cagtgttcct gcgccggttg cattcgattc ctgtttgtaa ttgtcctttt aacagcgatc
      541 gcgtatttcg tctcgctcag gcgcaatcac gaatgaataa cggtttggtt gatgcgagtg
      601 attttgatga cgagcgtaat ggctggcctg ttgaacaagt ctggaaagaa atgcataaac
      661 ttttgccatt ctcaccggat tcagtcgtca ctcatggtga tttctcactt gataacctta
      721 tttttgacga ggggaaatta ataggttgta ttgatgttgg acgagtcgga atcgcagacc
      781 gataccagga tcttgccatc ctatggaact gcctcggtga gttttctcct tcattacaga
      841 aacggctttt tcaaaaatat ggtattgata atcctgatat gaataaattg cagtttcatt
      901 tgatgctcga tgagtttttc taatcagaat tggttaattg gttgtaacat tattcagatt
      961 gggcttgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     1021 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     1081 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     1141 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     1201 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     1261 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     1321 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     1381 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     1441 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     1501 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     1561 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     1621 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     1681 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     1741 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt
     1801 gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag
     1861 gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa
     1921 tgcagctggc acgacaggtt tcccgatact tctaatgctg tgacatcact cacaggattc
     1981 ctgcaagtaa tgcgacattg gatactttgg gattccttta tagtcagtgg gttatccgaa
     2041 tttttgcctt gttcgttgct acattggcta aatcagaact tcggcagctc gtcctggaaa
     2101 gcgggcagtg agcgcaacgc aattaatgtg agttagctca ctcattaggc accccaggct
     2161 ttacacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata acaatttcac
     2221 acaggaaaca gctatgacca tgattacgcc aagcttgcat gcctgcaggt cgactctaga
     2281 ggatccccgg gtaccgagct cgaattcact ggccgtcgtt ttacaacgtc gtgactggga
     2341 aaaccctggc gttacccaac ttaatcgcct tgcagcacat ccccctttcg ccagctggcg
     2401 taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga
     2461 atggcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatatg
     2521 gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagcccc gacacccgcc
     2581 aacaccctcg gtcgaattcg gactgggatt ctccaatatt ccttagcgct tccaataagc
     2641 tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt
     2701 ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cga
//
