LOCUS       sequence-171182-        6970 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor vector for 3' FLAG tag of human SAP130.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6970)
  AUTHORS   Savic D, Partridge EC, Newberry KM, Smith SB, Meadows SK, Roberts 
            BS, Mackiewicz M, Mendenhall EM, Myers RM
  TITLE     CETCh-seq: CRISPR epitope tagging ChIP-seq of DNA-binding proteins.
  JOURNAL   Genome Res. 2015 Sep 9.
  PUBMED    26355004
REFERENCE   2  (bases 1 to 6970)
  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: "Genome Res.
            2015 Sep 9."
COMMENT     Sequence Label: pFETCh_SAP130
FEATURES             Location/Qualifiers
     source          1..6970
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      14..602
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     503..522
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     756..773
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    1073..1094
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1109..1114,1115..1132,1133..1139)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1147..1163
                     /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     1152..1174
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1171..1187
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1171..1187
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        1364..1566
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1517..1537
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     1563..1587
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             1663..2793
                     /codon_start=1
                     /product="herpes simplex virus thymidine kinase"
                     /label=HSV TK
                     /note="confers sensitivity to nucleoside analogs such as 
                     acyclovir or ganciclovir"
                     /translation="MASYPCHQHASAFDQAARSRGHSNRRTALRPRRQQEATEVRLEQK
                     MPTLLRVYIDGPHGMGKTTTTQLLVALGSRDDIVYVPEPMTYWQVLGASETIANIYTTQ
                     HRLDQGEISAGDAAVVMTSAQITMGMPYAVTDAVLAPHIGGEAGSSHAPPPALTLIFDR
                     HPIAALLCYPAARYLMGSMTPQAVLAFVALIPPTLPGTNIVLGALPEDRHIDRLAKRQR
                     PGERLDLAMLAAIRRVYGLLANTVRYLQGGGSWREDWGQLSGTAVPPQGAEPQSNAGPR
                     PHIGDTLFTLFRAPELLAPNGDLYNVFAWALDVLAKRLRPMHVFILDYDQSPAGCRDAL
                     LQLTSGMVQTHVTTPGSIPTICDLARTFAREMGEAN"
     CDS             3827..3850
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             3854..3877
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             3881..3904
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             3920..3976
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             3989..4783
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4043..4062)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4653..4672
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5554..5570)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5563..5585)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             complement(5789..6604)
                     /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="MSHIQRETSCSKPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNILQFHLMLDEFF"
     primer_bind     6512..6531
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     6596..6615
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
ORIGIN
        1 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
       61 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
      121 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
      181 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
      241 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
      301 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
      361 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
      421 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
      481 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
      541 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
      601 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
      661 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag
      721 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg
      781 aagaagctcg cacattcagc agcgtttttc agcgcgtttt cgatcaacgt ttcaatgttg
      841 gtatcaacac caggtttaac tttgaactta tcggcactga cggttactga ttttgaactt
      901 ttgctttgcc acggaacggt ctgcgttgtc gggaagatgc gtgatctgat ccttcaactc
      961 agcaaaagtt cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca
     1021 gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga
     1081 gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt
     1141 gtggaattgt gagcggataa caatttcaca caggaaacag ctatgaccat gattacgcca
     1201 agcttgcatg cctgcaggtc gactctagag gatccccggg taccgagctc gaattagctt
     1261 gcatgcctgc aggtcaattc cctggcatta tgcccagtac atgaccttat gggactttcc
     1321 tacttggcag tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca
     1381 gtacatcaat gggcgtggat agcggtttga ctcacgggga tttcaagtct ccaccccatt
     1441 gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa atgtcgtaac
     1501 aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt ctatataagc
     1561 agagctcgtt tagtgaaccg tcagatcgcc tggagacgcc atccacgctg ttttgacctc
     1621 cataggagac accgggaccg atccagcctg gggatcccga ccatggcttc gtacccctgc
     1681 catcaacacg cgtctgcgtt cgaccaggct gcgcgttctc gcggccatag caaccgacgt
     1741 acggcgttgc gccctcgccg gcagcaagaa gccacggaag tccgcctgga gcagaaaatg
     1801 cccacgctac tgcgggttta tatagacggt cctcacggga tggggaaaac caccaccacg
     1861 caactgctgg tggccctggg ttcgcgcgac gatatcgtct acgtacccga gccgatgact
     1921 tactggcagg tgctgggggc ttccgagaca atcgcgaaca tctacaccac acaacaccgc
     1981 ctcgaccagg gtgagatatc ggccggggac gcggcggtgg taatgacaag cgcccagata
     2041 acaatgggca tgccttatgc cgtgaccgac gccgttctgg ctcctcatat cgggggggag
     2101 gctgggagct cacatgcccc gcccccggcc ctcaccctca tcttcgaccg ccatcccatc
     2161 gccgccctcc tgtgctaccc ggccgcgcga taccttatgg gcagcatgac cccccaggcc
     2221 gtgctggcgt tcgtggccct catcccgccg accttgcccg gcacaaacat cgtgttgggg
     2281 gcccttccgg aggacagaca catcgaccgc ctggccaaac gccagcgccc cggcgagcgg
     2341 cttgacctgg ctatgctggc cgcgattcgc cgcgtttacg ggctgcttgc caatacggtg
     2401 cggtatctgc agggcggcgg gtcgtggcgg gaggattggg gacagctttc ggggacggcc
     2461 gtgccgcccc agggtgccga gccccagagc aacgcgggcc cacgacccca tatcggggac
     2521 acgttattta ccctgtttcg ggcccccgag ttgctggccc ccaacggcga cctgtacaac
     2581 gtgtttgcct gggccttgga cgtcttggcc aaacgcctcc gtcccatgca cgtctttatc
     2641 ctggattacg accaatcgcc cgccggctgc cgggacgccc tgctgcaact tacctccggg
     2701 atggtccaga cccacgtcac cacccccggc tccataccga cgatctgcga cctggcgcgc
     2761 acgtttgccc gggagatggg ggaggctaac tgaggatccc acgtcactat tgtatactct
     2821 atattatact ctatgttata ctctgtaatc ctactcaata aacgtgtcac gcctgtgaaa
     2881 ccgtactaag tctcccgtgt attcttatca ccatcaggtg acatcctcgc ccaggctgtc
     2941 aatcatgccg gtatcgattc cagtagcacc ggccccacgc tgacaaccca ctcttgcagc
     3001 gttagcagcg cccctcttaa caagccgacc cccaccagcg tcgcggttac taacactcct
     3061 ctccccgacc tgcagcccag ctgcgcccag cccctttact gtttttaaac cttagtatct
     3121 tagtccttag gagaaaggac gttgtcaccc aaacaactct tagacagttt acaaaggtga
     3181 ttgaaggtgg gttcatttgc ctggggttga catagaaaaa tatttccgag cttatgatac
     3241 cctccctttt gtgtgttgca gacgaatcta gaacatgatg tctatgaaag acttactaac
     3301 ctgcaggaag ggattatccc aaagaaaaaa gcagcaacag atgatgatct ccaccgaata
     3361 aacgaactga tacaggtttg agtgcgcccc ttcccttctg atcatctact ttatgaagtt
     3421 gctttctgta aagccctact tctcccgttt ccccccgtct ccctcatatc ttcagtaaaa
     3481 acatacgtgc ttcagcctgc actcctgaga aacaaaaaga cattttctct gttgtggaag
     3541 cctcaacatg gcctctgccc aaaggcaagc agttttttaa aagctgtact atcctcatgg
     3601 tcactgattg atgatgcaat gtcttctgtt tcagggaaat atgcagaggt gtaaacttgt
     3661 gatggatcaa atcagtgaag ccagagactc catgcttaag gttttagatc ataaagaccg
     3721 tgtcctgaag ctgcttaaca agaacgggac tgtcaaaaaa gtgtccaaat tgaagcgaaa
     3781 ggaaaaagtc gggagcggag gaggttccgg tggaggtggt tctggagatt acaaggatga
     3841 cgacgataag ggcgattaca aggatgacga cgataaggga gattacaagg atgacgacga
     3901 taaggtttca ggaagcggag ctactaactt cagcctgctg aagcaggctg gagacgtgga
     3961 ggagaaccct ggacctggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc
     4021 tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg
     4081 ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac
     4141 cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc
     4201 cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg
     4261 gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga
     4321 gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg
     4381 cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg
     4441 tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt
     4501 cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc
     4561 ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg
     4621 gctgggtgtg gccgaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga
     4681 gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc
     4741 gcagcgcatc gccttctatc gccttcttga cgagttcttc tgattcgaac atcatcagga
     4801 gattggaagc aaatttatga agaatgatgg tgggggtggg gggagggttt tggttttttc
     4861 caaagtggaa cattgaaata aaggaagtgt tccttagttc ccgtgtgaaa gcagaggaac
     4921 ccatgacatc caagggcgtg aaaggatcag agctgactgg acatagtgag ctgccttctt
     4981 gcgttcgggt gcacccctgt taaacctgat ctgtgtcata agtgactccg gatgcatcag
     5041 tgtccaccag ttggaagcaa tgacaaggat ggctggctgg tgtttttcag ccttccggtt
     5101 tatagactgt atttatctag tggattcctg caggccccat actgagcctg gactgaaagt
     5161 atccactcgg accatctgtt atctctctac actgaaaata aaacctcttc cacccacccc
     5221 attcggttct tctgcctgac cttcaaatgc ccatgttggc cttttacagc agtgccacgg
     5281 caccaagcga gctgccacat ctcacactct aaagggtttg aactattagt tcttgtcatt
     5341 ttttaaaaaa aaccattccc aagtgaaatt gttatatcgt ctgtcttgcg tgtgtcagaa
     5401 ctgggttttt gtggaggttc agagcaggca acaccataag ttgctctcag atccttgttc
     5461 tgaagtacat tcttggttat ctgtacttct gtagctggtg ccttggaaag tcctctccac
     5521 tgactgtagc ctccaattca ctggccgtcg ttcactggcc gtcgttttac aacgtcgtga
     5581 ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag
     5641 ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa
     5701 tggcgaatgc gatttattca acaaagccgc cgtcccgtca agtcagcgta atgctctgcc
     5761 agtgttacaa ccaattaacc aattctgatt agaaaaactc atcgagcatc aaatgaaact
     5821 gcaatatatt catatcagga ttatcaatac catatttttg aaaaagccgt ttctgtaatg
     5881 aaggagaaaa ctcaccgagg cagttccata ggatggcaag atcctggtat cggtctgcga
     5941 ttccgactcg tccaacatca atacaaccta ttaatttccc ctcgtcaaaa ataaggttat
     6001 caagtgagaa atcaccatga gtgacgactg aatccggtga gaatggcaaa agcttatgca
     6061 tttctttcca gacttgttca acaggccagc cattacgctc gtcatcaaaa tcactcgcat
     6121 caaccaaacc gttattcatt cgtgattgcg cctgagcgag acgaaatacg cgatcgctgt
     6181 taaaaggaca attacaaaca ggaatcgaat gcaaccggcg caggaacact gccagcgcat
     6241 caacaatatt ttcacctgaa tcaggatatt cttctaatac ctggaatgct gttttcccgg
     6301 ggatcgcagt ggtgagtaac catgcatcat caggagtacg gataaaatgc ttgatggtcg
     6361 gaagaggcat aaattccgtc agccagttta gtctgaccat ctcatctgta acatcattgg
     6421 caacgctacc tttgccatgt ttcagaaaca actctggcgc atcgggcttc ccatacaatc
     6481 gatagattgt cgcacctgat tgcccgacat tatcgcgagc ccatttatac ccatataaat
     6541 cagcatccat gttggaattt aatcgcggct tcgagcaaga cgtttcccgt tgaatatggc
     6601 tcataacacc ccttgtatta ctgtttatgt aagcagacag ttttattgtt catgatgata
     6661 tatttttatc ttgtgcaatg taacatcaga gattttgaga cacaacgtgg ctttgttgaa
     6721 taaatcgaac ttttgctgag ttgaaggatc agatcacgca tcttcccgac aacgcagacc
     6781 gttccgtggc aaagcaaaag ttcaaaatca ccaactggtc cacctacaac aaagctctca
     6841 tcaaccgtgg ctccctcact ttctggctgg atgatggggc gattcaggcc tggtatgagt
     6901 cagcaacacc ttcttcacga ggcagacctc tcgacggagt tccactgagc gtcagacccc
     6961 gtagaaaaga
//
