LOCUS       sequence-256203-        2961 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  SV40 promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2961)
  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 2961)
  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: S6_BstBI_SV40_SwaI
FEATURES             Location/Qualifiers
     source          1..2961
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..17
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        99..309
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        121..317
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      168..303
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(407..423)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(407..423)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(420..442)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    431..447
                     /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(455..461,462..479,480..485))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    500..521
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(638..655)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(809..1397)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(889..908)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(1568..2383)
                     /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(1568..2383)
                     /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     2291..2310
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(2384..2488)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2556..2574
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2612..2634)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2734..2753
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     join(2947..2961,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(2961,1..17)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
ORIGIN
        1 gtaaaacgac ggccagtgaa ttgacgcgta ttgggatgga acttcgaatg catctcaatt
       61 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
      121 tgcgtctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
      181 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
      241 aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag
      301 gcctaggctt ttgcaaaaag ctttctagag gtaccaccgg agcttattta aatacacaat
      361 taattaagga acatcccaat ggcgcgccga gcttggctcg agcatggtca tagctgtttc
      421 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
      481 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
      541 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
      601 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
      661 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
      721 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
      781 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
      841 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
      901 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
      961 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     1021 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     1081 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     1141 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     1201 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     1261 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     1321 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     1381 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     1441 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     1501 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     1561 ctgacagtta gaaaaactca tcgagcatca aatgaaactg caatttattc atatcaggat
     1621 tatcaatacc atatttttga aaaagccgtt tctgtaatga aggagaaaac tcaccgaggc
     1681 agttccatag gatggcaaga tcctggtatc ggtctgcgat tccgactcgt ccaacatcaa
     1741 tacaacctat taatttcccc tcgtcaaaaa taaggttatc aagtgagaaa tcaccatgag
     1801 tgacgactga atccggtgag aatggcaaaa gtttatgcat ttctttccag acttgttcaa
     1861 caggccagcc attacgctcg tcatcaaaat cactcgcatc aaccaaaccg ttattcattc
     1921 gtgattgcgc ctgagcgaga cgaaatacgc gatcgctgtt aaaaggacaa ttacaaacag
     1981 gaatcgaatg caaccggcgc aggaacactg ccagcgcatc aacaatattt tcacctgaat
     2041 caggatattc ttctaatacc tggaatgctg ttttcccagg gatcgcagtg gtgagtaacc
     2101 atgcatcatc aggagtacgg ataaaatgct tgatggtcgg aagaggcata aattccgtca
     2161 gccagtttag tctgaccatc tcatctgtaa catcattggc aacgctacct ttgccatgtt
     2221 tcagaaacaa ctctggcgca tcgggcttcc catacaatcg atagattgtc gcacctgatt
     2281 gcccgacatt atcgcgagcc catttatacc catataaatc agcatccatg ttggaattta
     2341 atcgcggcct agagcaagac gtttcccgtt gaatatggct catactcttc ctttttcaat
     2401 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     2461 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct
     2521 aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc
     2581 gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg
     2641 tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg
     2701 gtgttggcgg gtgtcggggc tggcttaact atgcggcatc agagcagatt gtactgagag
     2761 tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc
     2821 gccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc
     2881 tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg gtaacgccag
     2941 ggttttccca gtcacgacgt t
//
