LOCUS       sequence-435075-        3161 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  golden gate cloning.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3161)
  AUTHORS   Delhaye L, Moschonas GD, Fijalkowska D, Verhee A, De Sutter D, Van 
            de Steene T, De Meyer M, Grzesik H, Van Moortel L, De Bosscher K, 
            Jacobs T, Eyckerman S
  TITLE     Leveraging a self-cleaving peptide for tailored control in proximity
            labeling proteomics.
  JOURNAL   Cell Rep Methods. 2024 Jul 15;4(7):100818. doi: 
            10.1016/j.crmeth.2024.100818. Epub 2024 Jul 9.
  PUBMED    38986614
REFERENCE   2  (bases 1 to 3161)
  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 Rep 
            Methods."; date: "2024-07-15"; volume: "4(7):100818. doi"; pages: " 
            10.1016/j.crmeth.2024.100818. Epub 2024 Jul 9"
COMMENT     Sequence Label: pEN-X-hPGK-BlastR-Z
FEATURES             Location/Qualifiers
     source          1..3161
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     297..315
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     CDS             387..785
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     primer_bind     complement(893..909)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(893..909)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(906..928)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     rep_origin      complement(1081..1669)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1161..1180)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(1805..2620)
                     /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(1805..2620)
                     /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     2528..2547
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2612..2631
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     2741..2757
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2741..2757
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     terminator      2758..2804
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     2918..2940
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2932..2949
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2933..2949
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        join(3029..3161,1..378)
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
ORIGIN
        1 acccggatct tcgccgctac ccttgtgggc cccccggcga cgcttcctgc tccgccccta
       61 agtcgggaag gttccttgcg gttcgcggcg tgccggacgt gacaaacgga agccgcacgc
      121 ctcactagta ccctcgcaga cggacagcgc cagggagcaa tggcagcgcg ccgaccgcga
      181 tgggctgtgg ccaatagcgg ctgctcagca gggcgcgccg agagcagcgg ccgggaaggg
      241 gcggtgcggg aggcggggtg tggggcggta gtgtgggccc tgttcctgcc cgcgcggtgt
      301 tccgcattct gcaagcctcc ggagcgcacg tcggcagtcg gctccctcgt tgaccgaatc
      361 accgacctct ctccccagaa cagaccatgg ccaagccttt gtctcaagaa gaatccaccc
      421 tcattgaaag agcaacggct acaatcaaca gcatccccat ctctgaagac tacagcgtcg
      481 ccagcgcagc tctctctagc gacggccgca tcttcactgg tgtcaatgta tatcatttta
      541 ctgggggacc ttgtgcagaa ctcgtggtgc tgggcactgc tgctgctgcg gcagctggca
      601 acctgacttg tatcgtcgcg atcggaaatg agaacagggg catcttgagc ccctgcggac
      661 ggtgccgaca ggtgcttctc gatctgcatc ctgggatcaa agccatagtg aaggacagtg
      721 atggacagcc gacggcagtt gggattcgtg aattgctgcc ctctggttat gtgtgggagg
      781 gctaaggctc taaggttaag tcgccctcgg tatcagatga gacgcgtata gtcttcaggc
      841 taggtggagg ctcagtgatg ataagtctgc gatggtggat gcatgtgtca tggtcatagc
      901 tgtttcctgt gtgaaattgt tatccgctca gagggcacaa tcctattccg cgctatccga
      961 caatctccaa gacattaggt ggagttcagt tcggcgtatg gcatatgtcg ctggaaagaa
     1021 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     1081 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     1141 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     1201 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     1261 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     1321 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     1381 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     1441 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     1501 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     1561 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     1621 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     1681 gatcttttct acggggtctg acgctctatt caacaaagcc gccgtcccgt caagtcagcg
     1741 taaatgggta gggggcttca aatcgtccgc tctgccagtg ttacaaccaa ttaacaaatt
     1801 ctgattagaa aaactcatcg agcatcaaat gaaactgcaa tttattcata tcaggattat
     1861 caataccata tttttgaaaa agccgtttct gtaatgaagg agaaaactca ccgaggcagt
     1921 tccataggat ggcaagatcc tggtatcggt ctgcgattcc gactcgtcca acatcaatac
     1981 aacctattaa tttcccctcg tcaaaaataa ggttatcaag tgagaaatca ccatgagtga
     2041 cgactgaatc cggtgagaat ggcaaaagct tatgcatttc tttccagact tgttcaacag
     2101 gccagccatt acgctcgtca tcaaaatcac tcgcatcaac caaaccgtta ttcattcgtg
     2161 attgcgcctg agccagacga aatacgcgat cgctgttaaa aggacaatta caaacaggaa
     2221 tcgaatgcaa ccggcgcagg aacactgcca gcgcatcaac aatattttca cctgaatcag
     2281 gatattcttc taatacctgg aatgctgttt tcccggggat cgcagtggtg agtaaccatg
     2341 catcatcagg agtacggata aaatgcttga tggtcggaag aggcataaat tccgtcagcc
     2401 agtttagtct gaccatctca tctgtaacat cattggcaac gctacctttg ccatgtttca
     2461 gaaacaactc tggcgcatcg ggcttcccat acaatcgata gattgtcgca cctgattgcc
     2521 cgacattatc gcgagcccat ttatacccat ataaatcagc atccatgttg gaatttaatc
     2581 gcggcctcga gcaagacgtt tcccgttgaa tatggctcat aacacccctt gtattactgt
     2641 ttatgtaagc agacagtttt attgttcatg atgatatatt tttatcttgt gcaatgtaac
     2701 atcagagatt ttgagacaca acgtggcttt cccccgccgc tctagaacta gtggatccaa
     2761 ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt ttatctgttg tttgtcgcat
     2821 tatagcagac gtccaggttg ggatacctga aacaaaaccc atcgtacggc caaggaagtc
     2881 tccaataact gtgatccacc acaagcgcca gggttttccc agtcacgacg ttgtaaaacg
     2941 acggccagtc atgcataatc cgcacgcatc tggaataagg aagtgccatt ccgcctgacc
     3001 tgaagacgtg tgacgtctcc gcaaacctgg ggttggggtt gcgccttttc caaggcagcc
     3061 ctgggtttgc gcagggacgc ggctgctctg ggcgtggttc cgggaaacgc agcggcgccg
     3121 accctgggac tcgcacattc ttcacgtccg ttcgcagcgt c
//
