LOCUS       sequence-463097-        5529 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway pDONR vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5529)
  AUTHORS   Marohl T, Atkins KA, Wang L, Janes KA
  TITLE     PCSK5M452I is a recessive hypomorph exclusive to MCF10DCIS.com 
            cells.
  JOURNAL   Mol Cancer Res. 2025 Oct 28. doi: 10.1158/1541-7786.MCR-25-0211.
  PUBMED    41148168
REFERENCE   2  (bases 1 to 5529)
  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: "Mol Cancer 
            Res. 2025 Oct 28. doi: 10.1158/1541-7786.MCR-25-0211."
COMMENT     Sequence Label: pDONR223 PCSK5(M452I)
FEATURES             Location/Qualifiers
     source          1..5529
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     92..109
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(268..295)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(387..473)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     522..544
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     536..553
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     537..553
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    569..668
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(3408..3507)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(3524..3543)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3525..3543)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3548..3564)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3548..3564)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             3995..4786
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     rep_origin      4879..5467
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5368..5387
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
       61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
      121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
      181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc
      241 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta
      301 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc
      361 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa
      421 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg
      481 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa
      541 aacgacggcc agtcttaagc tcgggcccca aataatgatt ttattttgac tgatagtgac
      601 ctgttcgttg caacaaattg atgagcaatg cttttttata atgccaactt tgtacaaaaa
      661 agttggcatg ggctggggga gccgctgctg ctgcccggga cgtttggacc tgctgtgcgt
      721 gctggcgctg ctcgggggct gcctgctccc cgtgtgtcgg acgcgcgtct acaccaacca
      781 ctgggcagtc aaaatcgccg ggggcttccc ggaggccaac cgtatcgcca gcaagtacgg
      841 attcatcaac ataggacaga taggggccct gaaggactac taccacttct accatagcag
      901 gacgattaaa aggtcagtta tctcgagcag agggacccac agtttcattt caatggaacc
      961 aaaggtggaa tggatccaac agcaagtggt aaaaaagcgg acaaagaggg attatgactt
     1021 cagtcgtgcc cagtctacct atttcaatga tcccaagtgg cccagcatgt ggtatatgca
     1081 ctgcagtgac aatacacatc cctgccagtc tgacatgaat atcgaaggag cctggaagag
     1141 aggctacacg ggaaagaaca ttgtggtcac tatcctggat gacggaattg agagaaccca
     1201 tccagatctg atgcaaaact acgatgctct ggcaagttgc gacgtgaatg ggaatgactt
     1261 ggacccaatg cctcgttatg atgcaagcaa cgagaacaag catgggactc gctgtgctgg
     1321 agaagtggca gccgctgcaa acaattcgca ctgcacagtc ggaattgctt tcaacgccaa
     1381 gatcggagga gtgcgaatgc tggacggaga tgtcacggac atggttgaag caaaatcagt
     1441 tagcttcaac ccccagcacg tgcacattta cagcgccagc tggggcccgg atgatgatgg
     1501 caagactgtg gacggaccag cccccctcac ccggcaagcc tttgaaaacg gcgttagaat
     1561 ggggcggaga ggcctcggct ctgtgtttgt ttgggcatct ggaaatggtg gaaggagcaa
     1621 agaccactgc tcctgtgatg gctacaccaa cagcatctac accatctcca tcagcagcac
     1681 tgcagaaagc ggaaagaaac cttggtacct ggaagagtgt tcatccacgc tggccacaac
     1741 ctacagcagc ggggagtcct acgataagaa aatcatcact acagatctga ggcagcgttg
     1801 cacggacaac cacactggga cgtcagcctc agcccccatg gctgcaggca tcattgcgct
     1861 ggccctggaa gccaatccgt ttctgacctg gagagacgta cagcatgtta ttgtcaggac
     1921 ttcccgtgcg ggacatttga acgctaatga ctggaaaacc aatgctgctg gttttaaggt
     1981 gagccatctt tatggatttg gactgatgga cgcagaagcc attgtgatgg aggcagagaa
     2041 gtggaccacc gttccccggc agcacgtgtg tgtggagagc acagaccgac aaatcaagac
     2101 aatccgccct aacagtgcag tgcgctccat ctacaaagct tcaggctgct cggataaccc
     2161 caaccgccat gtcaactacc tggagcacgt cgttgtgcgc atcaccatca cccaccccag
     2221 gagaggagac ctggccatct acctgacctc gccctctgga actaggtctc agcttttggc
     2281 caacaggcta tttgatcact ccatggaagg attcaaaaac tgggagttca tgaccattca
     2341 ttgctgggga gaaagagctg ctggtgactg ggtccttgaa gtttatgata ctccctctca
     2401 gctaaggaac tttaagactc caggtaaatt gaaagaatgg tctttggtcc tctacggcac
     2461 ctccgtgcag ccatattcac caaccaatga atttccgaaa gtggaacggt tccgctatag
     2521 ccgagttgaa gaccccacag acgactatgg cacagaggat tatgcaggtc cctgcgaccc
     2581 tgagtgcagt gaggttggct gtgacgggcc aggaccagac cactgcaatg actgtttgca
     2641 ctactactac aagctgaaaa acaataccag gatctgtgtc tccagctgcc cccctggcca
     2701 ctaccacgcc gacaagaagc gctgcaggaa gtgtgccccc aactgtgagt cctgctttgg
     2761 gagccatggt gaccaatgca tgtcctgcaa atatggatac tttctgaatg aagaaaccaa
     2821 cagctgtgtt actcactgcc ctgatgggtc atatcaggat accaagaaaa atctttgccg
     2881 gaaatgcagt gaaaactgca agacatgtac tgaattccat aactgtacag aatgtaggga
     2941 tgggttaagc ctgcagggat cccggtgctc tgtctcctgt gaagatggac ggtatttcaa
     3001 cggccaggac tgccagccct gccaccgctt ctgcgccact tgtgctgggg caggagctga
     3061 tgggtgcatt aactgcacag agggctactt catggaggat gggagatgcg tgcagagctg
     3121 tagtatcagc tattactttg accactcttc agagaatgga tacaaatcct gcaaaaaatg
     3181 tgatatcagt tgtttgacgt gcaatggccc aggattcaag aactgtacaa gctgccctag
     3241 tgggtatctc ttagacttag gaatgtgtca aatgggagcc atttgcaagg atgcaacgga
     3301 agagtcctgg gcggaaggag gcttctgtat gcttgtgaaa aagaacaatc tgtgccaacg
     3361 gaaggttctt caacaacttt gctgcaaaac atgtacattt caaggctgcc caactttctt
     3421 gtacaaagtt ggcattataa gaaagcattg cttatcaatt tgttgcaacg aacaggtcac
     3481 tatcagtcaa aataaaatca ttatttgcca tccagctgat atcccctata gtgagtcgta
     3541 ttacatggtc atagctgttt cctggcagct ctggcccgtg tctcaaaatc tctgatgtta
     3601 cattgcacaa gataaaaata tatcatcatg cctcctctag accagccagg acagaaatgc
     3661 ctcgacttcg ctgctaccca aggttgccgg gtgacgcaca ccgtggaaac ggatgaaggc
     3721 acgaacccag tggacataag cctgttcggt tcgtaagctg taatgcaagt agcgtatgcg
     3781 ctcacgcaac tggtccagaa ccttgaccga acgcagcggt ggtaacggcg cagtggcggt
     3841 tttcatggct tgttatgact gtttttttgg ggtacagtct atgcctcggg catccaagca
     3901 gcaagcgcgt tacgccgtgg gtcgatgttt gatgttatgg agcagcaacg atgttacgca
     3961 gcagggcagt cgccctaaaa caaagttaaa cattatgagg gaagcggtga tcgccgaagt
     4021 atcgactcaa ctatcagagg tagttggcgt catcgagcgc catctcgaac cgacgttgct
     4081 ggccgtacat ttgtacggct ccgcagtgga tggcggcctg aagccacaca gtgatattga
     4141 tttgctggtt acggtgaccg taaggcttga tgaaacaacg cggcgagctt tgatcaacga
     4201 ccttttggaa acttcggctt cccctggaga gagcgagatt ctccgcgctg tagaagtcac
     4261 cattgttgtg cacgacgaca tcattccgtg gcgttatcca gctaagcgcg aactgcaatt
     4321 tggagaatgg cagcgcaatg acattcttgc aggtatcttc gagccagcca cgatcgacat
     4381 tgatctggct atcttgctga caaaagcaag agaacatagc gttgccttgg taggtccagc
     4441 ggcggaggaa ctctttgatc cggttcctga acaggatcta tttgaggcgc taaatgaaac
     4501 cttaacgcta tggaactcgc cgcccgactg ggctggcgat gagcgaaatg tagtgcttac
     4561 gttgtcccgc atttggtaca gcgcagtaac cggcaaaatc gcgccgaagg atgtcgctgc
     4621 cgactgggca atggagcgcc tgccggccca gtatcagccc gtcatacttg aagctagaca
     4681 ggcttatctt ggacaagaag aagatcgctt ggcctcgcgc gcagatcagt tggaagaatt
     4741 tgtccactac gtgaaaggcg agatcaccaa ggtagtcggc aaataaccct cgagccaccc
     4801 atgaccaaaa tcccttaacg tgagttacgc gtcgttccac tgagcgtcag accccgtaga
     4861 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     4921 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     4981 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     5041 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     5101 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     5161 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     5221 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     5281 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     5341 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     5401 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     5461 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     5521 tcacatgtt
//
