LOCUS       sequence-432120-        2639 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Fragmid fragment: (N' terminus) SV40 Nuclear Localization Signal.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2639)
  AUTHORS   McGee AV, Liu YV, Griffith AL, Szegletes ZM, Wen B, Kraus C, Miller 
            NW, Steger RJ, Escude Velasco B, Bosch JA, Zirin JD, Viswanatha R, 
            Sontheimer EJ, Goodale A, Greene MA, Green TM, Doench JG
  TITLE     Modular vector assembly enables rapid assessment of emerging CRISPR 
            technologies.
  JOURNAL   Cell Genom. 2024 Mar 13;4(3):100519. doi: 
            10.1016/j.xgen.2024.100519.
  PUBMED    38484704
REFERENCE   2  (bases 1 to 2639)
  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: "Cell 
            Genom."; date: "2024-03-13"; volume: "4(3):100519. doi"; pages: " 
            10.1016/j.xgen.2024.100519"
COMMENT     Sequence Label: AA295
FEATURES             Location/Qualifiers
     source          1..2639
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      311..899
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     800..819
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1053..1070
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        join(1223..1228,1229..1246,1247..1253)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1261..1277
                     /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     1266..1288
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1285..1301
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1285..1301
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             1297..1704
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHAGLCSRRARDPMKTAAHLSLLLRHHGGGPTSSSRCI
                     REVPSSNSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGE
                     WRLMRYFLLTHLCGISHRIWCTLSTICSDAA"
     CDS             complement(1372..1392)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     regulatory      1395..1404
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(1464..1486)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(1680..1699)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1799..1821
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1952..1971)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             join(1963..2639,1..133)
                     /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(2030..2049)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
ORIGIN
        1 tcttgccatc ctatggaact gcctcggtga gttttctcct tcattacaga aacggctttt
       61 tcaaaaatat ggtattgata atcctgatat gaataaattg cagtttcatt tgatgctcga
      121 tgagtttttc taatcagaat tggttaattg gttgtaacat tattcagatt gggcttgatt
      181 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
      241 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
      301 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
      361 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
      421 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
      481 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
      541 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
      601 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
      661 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
      721 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
      781 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
      841 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
      901 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
      961 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg
     1021 ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag
     1081 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
     1141 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtaagttagc
     1201 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
     1261 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac accaagcttg
     1321 catgcaggcc tctgcagtcg acgggcccgg gatccgatga agactgctgc ccacctttct
     1381 cttcttctta ggcaccatgg tggcggtcct acgtcttcat ctagatgtat tcgcgaggta
     1441 ccgagctcga attctctggc cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt
     1501 acccaactta atcgccttgc agcacatccc cctttcgcca gctggcgtaa tagcgaagag
     1561 gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg gcgcctgatg
     1621 cggtattttc tccttacgca tctgtgcggt atttcacacc gcatatggtg cactctcagt
     1681 acaatctgct ctgatgccgc atagttaagc cagccccgac acccgccaac acccgctgac
     1741 gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc
     1801 gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgat gcagctctgg
     1861 cccgtgtctc aaaatctctg atgttacatt gcacaagata aaaatatatc atcatgaaca
     1921 ataaaactgt ctgcttacat aaacagtaat acaaggggtg ttatgagcca tattcaacgg
     1981 gaaacgtcga ggccgcgatt aaattccaac atggatgctg atttatatgg gtataaatgg
     2041 gctcgcgata atgtcgggca atcaggtgcg acaatctatc gcttgtatgg gaagcccgat
     2101 gcgccagagt tgtttctgaa acatggcaaa ggtagcgttg ccaatgatgt tacagatgag
     2161 atggtcagac taaactggct gacggaattt atgcctcttc cgaccatcaa gcattttatc
     2221 cgtactcctg atgatgcatg gttactcacc actgcgatcc ccggaaaaac agcattccag
     2281 gtattagaag aatatcctga ttcaggtgaa aatattgttg atgcgctggc agtgttcctg
     2341 cgccggttgc attcgattcc tgtttgtaat tgtcctttta acagcgatcg cgtatttcgt
     2401 ctcgctcagg cgcaatcacg aatgaataac ggtttggttg atgcgagtga ttttgatgac
     2461 gagcgtaatg gctggcctgt tgaacaagtc tggaaagaaa tgcataaact tttgccattc
     2521 tcaccggatt cagtcgtcac tcatggtgat ttctcacttg ataaccttat ttttgacgag
     2581 gggaaattaa taggttgtat tgatgttgga cgagtcggaa tcgcagaccg ataccagga
//
