LOCUS       sequence-161158-        2891 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  5' entry vector containing human phosphoglycerate kinase (hPGK) 
            promoter for ubiquitous expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2891)
  AUTHORS   Fowler DK, Stewart S, Seredick S, Eisen JS, Stankunas K, Washbourne 
            P
  TITLE     A MultiSite Gateway Toolkit for Rapid Cloning of Vertebrate 
            Expression Constructs with Diverse Research Applications.
  JOURNAL   PLoS One. 2016 Aug 8;11(8):e0159277. doi: 
            10.1371/journal.pone.0159277. eCollection 2016.
  PUBMED    27500400
REFERENCE   2  (bases 1 to 2891)
  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: "PLoS One.";
            date: "2016-08-8"; volume: "11(8):e0159277. doi"; pages: " 
            10.1371/journal.pone.0159277. eCollection 2016"
COMMENT     Sequence Label: p5E-hPGK
FEATURES             Location/Qualifiers
     source          1..2891
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(63..651)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(143..162)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(798..1607)
                     /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     1521..1540
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1599..1618
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     1720..1736
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1720..1736
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     1741..1760
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1741..1759
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1775..1870
                     /gene="mutant version of attL"
                     /label=attL4
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        1901..2411
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     2330..2348
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     protein_bind    2467..2590
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2783..2800)
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
       61 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
      121 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
      181 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
      241 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
      301 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
      361 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
      421 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
      481 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
      541 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
      601 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
      661 ttgatctttt ctacggggtc tgacgctcag tggaacgacg cgtaactcac gttaagggat
      721 tttggtcatg agcttgcgcc gtcccgtcaa gtcagcgtaa tgctctgcca gtgttacaac
      781 caattaacca attctgatta gaaaaactca tcgagcatca aatgaaactg caatttattc
      841 atatcaggat tatcaatacc atatttttga aaaagccgtt tctgtaatga aggagaaaac
      901 tcaccgaggc agttccatag gatggcaaga tcctggtatc ggtctgcgat tccgactcgt
      961 ccaacatcaa tacaacctat taatttcccc tcgtcaaaaa taaggttatc aagtgagaaa
     1021 tcaccatgag tgacgactga atccggtgag aatggcaaaa gtttatgcat ttctttccag
     1081 acttgttcaa caggccagcc attacgctcg tcatcaaaat cactcgcatc aaccaaaccg
     1141 ttattcattc gtgattgcgc ctgagcgaga cgaaatacgc gatcgctgtt aaaaggacaa
     1201 ttacaaacag gaatcgaatg caaccggcgc aggaacactg ccagcgcatc aacaatattt
     1261 tcacctgaat caggatattc ttctaatacc tggaatgctg tttttccggg gatcgcagtg
     1321 gtgagtaacc atgcatcatc aggagtacgg ataaaatgct tgatggtcgg aagaggcata
     1381 aattccgtca gccagtttag tctgaccatc tcatctgtaa catcattggc aacgctacct
     1441 ttgccatgtt tcagaaacaa ctctggcgca tcgggcttcc catacaagcg atagattgtc
     1501 gcacctgatt gcccgacatt atcgcgagcc catttatacc catataaatc agcatccatg
     1561 ttggaattta atcgcggcct cgacgtttcc cgttgaatat ggctcataac accccttgta
     1621 ttactgttta tgtaagcaga cagttttatt gttcatgatg atatattttt atcttgtgca
     1681 atgtaacatc agagattttg agacacgggc cagagctgcc aggaaacagc tatgaccatg
     1741 taatacgact cactataggg gatcatggag ctccaaataa tgattttatt ttgactgata
     1801 gtgacctgtt cgttgcaaca aattgataag caatgctttt ttataatgcc aactttgtat
     1861 agaaaagttg ggccggccct tcggcgcgat cgaattccac ggggttgggg ttgcgccttt
     1921 tccaaggcag ccctgggttt gcgcagggac gcggctgctc tgggcgtggt tccgggaaac
     1981 gcagcggcgc cgaccctggg tctcgcacat tcttcacgtc cgttcgcagc gtcacccgga
     2041 tcttcgccgc tacccttgtg ggccccccgg cgacgcttcc tgctccgccc ctaagtcggg
     2101 aaggttcctt gcggttcgcg gcgtgccgga cgtgacaaac ggaagccgca cgtctcacta
     2161 gtaccctcgc agacggacag cgccagggag caatggcagc gcgccgaccg cgatgggctg
     2221 tggccaatag cggctgctca gcagggcgcg ccgagagcag cggccgggaa ggggcggtgc
     2281 gggaggcggg gtgtggggcg gtagtgtggg ccctgttcct gcccgcgcgg tgttccgcat
     2341 tctgcaagcc tccggagcgc acgtcggcag tcggctccct cgttgaccga atcaccgacc
     2401 tctctcccca gggggatcca ccggttcgtc gactagtcca gtgtggtgga attctgcaga
     2461 tgcgcccaag tttgtacaaa aaagttgaac gagaaacgta aaatgatata aatatcaata
     2521 tattaaatta gattttgcat aaaaaacaga ctacataata ctgtaaaaca caacatatgc
     2581 agtcactatg aaccaactac ttagatggta ttagtgacct gtagaattat cgcggccgcc
     2641 ctgcagctct agcggtacgc gtattaattg cgttgcgctc actgcccgct ttccagtcgg
     2701 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     2761 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     2821 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     2881 acgcaggaaa g
//
