LOCUS       sequence-254531-        3063 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Zeo Resistance gene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3063)
  AUTHORS   Hazelbaker DZ, Beccard A, Angelini G, Mazzucato P, Messana A, Lam D,
            Eggan K, Barrett LE
  TITLE     A multiplexed gRNA piggyBac transposon system facilitates efficient 
            induction of CRISPRi and CRISPRa in human pluripotent stem cells.
  JOURNAL   Sci Rep. 2020 Jan 20;10(1):635. doi: 10.1038/s41598-020-57500-1.
  PUBMED    31959800
REFERENCE   2  (bases 1 to 3063)
  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: "Sci Rep."; 
            date: "2020-01-20"; volume: "10(1):635. doi"; pages: " 
            10.1038/s41598-020-57500-1"
COMMENT     Sequence Label: S7_SwaI_Zeo_PacI
FEATURES             Location/Qualifiers
     source          1..3063
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(27..49)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     149..168
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     362..384
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     376..393
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     377..393
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      442..451
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             448..822
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     primer_bind     complement(885..901)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(885..901)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(898..920)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    909..925
                     /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(933..939,940..957,958..963))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    978..999
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1116..1133)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1287..1875)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1367..1386)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2046..2861)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(2046..2861)
                     /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="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     2769..2788
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(2862..2966)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3034..3052
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga gacggtcaca
       61 gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt
      121 ggcgggtgtc ggggctggct taactatgcg gcatcagagc agattgtact gagagtgcac
      181 catatgcggt gtgaaatacc gcacagatgc gtaaggagaa aataccgcat caggcgccat
      241 tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta
      301 cgccagctgg cgaaaggggg atgtgctgca aggcgattaa gttgggtaac gccagggttt
      361 tcccagtcac gacgttgtaa aacgacggcc agtgaattga cgcgtattgg gattagaggt
      421 accaccggag cttatttaaa tgccaccatg gccaagttga ccagtgccgt tccggtgctc
      481 accgcgcgcg acgtcgccgg agcggtcgag ttctggaccg accggctcgg gttctcccgg
      541 gacttcgtgg aggacgactt cgccggtgtg gtccgggacg acgtgaccct gttcatcagc
      601 gcggtccagg accaggtggt gccggacaac accctggcct gggtgtgggt gcgcggcctg
      661 gacgagctgt acgccgagtg gtcggaggtc gtgtccacga acttccggga cgcctccggg
      721 ccggccatga ccgagatcgg cgagcagccg tgggggcggg agttcgccct gcgcgacccg
      781 gccggcaact gcgtgcactt cgtggccgag gagcaggact aattaattaa cacaaatacg
      841 cgtatctgca atcccaatgg cgcgccgagc ttggctcgag catggtcata gctgtttcct
      901 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
      961 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
     1021 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     1081 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     1141 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     1201 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     1261 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     1321 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     1381 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     1441 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     1501 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     1561 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     1621 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     1681 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     1741 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     1801 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     1861 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     1921 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     1981 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     2041 gacagttaga aaaactcatc gagcatcaaa tgaaactgca atttattcat atcaggatta
     2101 tcaataccat atttttgaaa aagccgtttc tgtaatgaag gagaaaactc accgaggcag
     2161 ttccatagga tggcaagatc ctggtatcgg tctgcgattc cgactcgtcc aacatcaata
     2221 caacctatta atttcccctc gtcaaaaata aggttatcaa gtgagaaatc accatgagtg
     2281 acgactgaat ccggtgagaa tggcaaaagt ttatgcattt ctttccagac ttgttcaaca
     2341 ggccagccat tacgctcgtc atcaaaatca ctcgcatcaa ccaaaccgtt attcattcgt
     2401 gattgcgcct gagcgagacg aaatacgcga tcgctgttaa aaggacaatt acaaacagga
     2461 atcgaatgca accggcgcag gaacactgcc agcgcatcaa caatattttc acctgaatca
     2521 ggatattctt ctaatacctg gaatgctgtt ttcccaggga tcgcagtggt gagtaaccat
     2581 gcatcatcag gagtacggat aaaatgcttg atggtcggaa gaggcataaa ttccgtcagc
     2641 cagtttagtc tgaccatctc atctgtaaca tcattggcaa cgctaccttt gccatgtttc
     2701 agaaacaact ctggcgcatc gggcttccca tacaatcgat agattgtcgc acctgattgc
     2761 ccgacattat cgcgagccca tttataccca tataaatcag catccatgtt ggaatttaat
     2821 cgcggcctag agcaagacgt ttcccgttga atatggctca tactcttcct ttttcaatat
     2881 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     2941 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa
     3001 gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt
     3061 ctc
//
