LOCUS       sequence-170774-        7154 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor vector for 3' FLAG tag of human HBP1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7154)
  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 7154)
  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_HBP1
FEATURES             Location/Qualifiers
     source          1..7154
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      48..636
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     537..556
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     790..807
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    1107..1128
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1143..1148,1149..1166,1167..1173)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1181..1197
                     /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     1186..1208
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1205..1221
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1205..1221
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        1398..1600
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1551..1571
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     1597..1621
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             1697..2827
                     /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             3953..3976
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             3980..4003
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             4007..4030
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             4046..4102
                     /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             4115..4909
                     /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(4169..4188)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4779..4798
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5772..5788)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5781..5803)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             complement(6007..6822)
                     /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     6730..6749
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     6814..6833
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
ORIGIN
        1 gagttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
       61 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
      121 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
      181 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
      241 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
      301 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
      361 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
      421 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
      481 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
      541 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
      601 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
      661 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
      721 tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg
      781 accgagcgca gcgagtcagt gagcgaggaa gcggaagaag ctcgcacatt cagcagcgtt
      841 tttcagcgcg ttttcgatca acgtttcaat gttggtatca acaccaggtt taactttgaa
      901 cttatcggca ctgacggtta ctgattttga acttttgctt tgccacggaa cggtctgcgt
      961 tgtcgggaag atgcgtgatc tgatccttca actcagcaaa agttcccaat acgcaaaccg
     1021 cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg
     1081 aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta ggcaccccag
     1141 gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt
     1201 cacacaggaa acagctatga ccatgattac gccaagcttg catgcctgca ggtcgactct
     1261 agaggatccc cgggtaccga gctcgaatta gcttgcatgc ctgcaggtca attccctggc
     1321 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
     1381 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
     1441 ttgactcacg gggatttcaa gtctccaccc cattgacgtc aatgggagtt tgttttggca
     1501 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg
     1561 cggtaggcgt gtacggtggg aggtctatat aagcagagct cgtttagtga accgtcagat
     1621 cgcctggaga cgccatccac gctgttttga cctccatagg agacaccggg accgatccag
     1681 cctggggatc ccgaccatgg cttcgtaccc ctgccatcaa cacgcgtctg cgttcgacca
     1741 ggctgcgcgt tctcgcggcc atagcaaccg acgtacggcg ttgcgccctc gccggcagca
     1801 agaagccacg gaagtccgcc tggagcagaa aatgcccacg ctactgcggg tttatataga
     1861 cggtcctcac gggatgggga aaaccaccac cacgcaactg ctggtggccc tgggttcgcg
     1921 cgacgatatc gtctacgtac ccgagccgat gacttactgg caggtgctgg gggcttccga
     1981 gacaatcgcg aacatctaca ccacacaaca ccgcctcgac cagggtgaga tatcggccgg
     2041 ggacgcggcg gtggtaatga caagcgccca gataacaatg ggcatgcctt atgccgtgac
     2101 cgacgccgtt ctggctcctc atatcggggg ggaggctggg agctcacatg ccccgccccc
     2161 ggccctcacc ctcatcttcg accgccatcc catcgccgcc ctcctgtgct acccggccgc
     2221 gcgatacctt atgggcagca tgacccccca ggccgtgctg gcgttcgtgg ccctcatccc
     2281 gccgaccttg cccggcacaa acatcgtgtt gggggccctt ccggaggaca gacacatcga
     2341 ccgcctggcc aaacgccagc gccccggcga gcggcttgac ctggctatgc tggccgcgat
     2401 tcgccgcgtt tacgggctgc ttgccaatac ggtgcggtat ctgcagggcg gcgggtcgtg
     2461 gcgggaggat tggggacagc tttcggggac ggccgtgccg ccccagggtg ccgagcccca
     2521 gagcaacgcg ggcccacgac cccatatcgg ggacacgtta tttaccctgt ttcgggcccc
     2581 cgagttgctg gcccccaacg gcgacctgta caacgtgttt gcctgggcct tggacgtctt
     2641 ggccaaacgc ctccgtccca tgcacgtctt tatcctggat tacgaccaat cgcccgccgg
     2701 ctgccgggac gccctgctgc aacttacctc cgggatggtc cagacccacg tcaccacccc
     2761 cggctccata ccgacgatct gcgacctggc gcgcacgttt gcccgggaga tgggggaggc
     2821 taactgagga tcccacgtca ctattgtata ctctatatta tactctatgt tatactctgt
     2881 aatcctactc aataaacgtg tcacgcctgt gaaaccgtac taagtctccc gtgtattctt
     2941 atcaccatca ggtgacatcc tcgcccaggc tgtcaatcat gccggtatcg attccagtag
     3001 caccggcccc acgctgacaa cccactcttg cagcgttagc agcgcccctc ttaacaagcc
     3061 gacccccacc agcgtcgcgg ttactaacac tcctctcccc gacctgcagc ccagctgtag
     3121 aaaatttctc taataatttt gggattgtct caacagcttt ttttccttaa ttggtgaggg
     3181 accaataagg ctggaggtgt tggggtatgt tagctgtcat tcatggggac ttgtgccatt
     3241 ggcacagaat ttctagattg aaaaatttca aggaggagaa gaaataggcc caggaatggt
     3301 ggcaaacaag tctatatgat aagctatctt ggatagactt gtttcttata acttttccat
     3361 tactaaacta gcatctttta ggttcagcgt ttttagatta attgcaaaaa aattacatgt
     3421 gatatagtgg ggtcaatttg aagtatcatt gaaagatgaa gggaagatat caatattttt
     3481 atagtaggag ggctggccaa agatgtttta aattcattgt gtacatgtgc ctgtttctta
     3541 gtaaaatgca ggtcatatct gtttaaattt ttcacaattt ctttaaaaat agtggtcagt
     3601 cagtagtcag atgagagggt ggctgaactg aattttggca gttcttggtt aatttagaat
     3661 ttcatgtgtt cggacttttc ctgtatccct tttctgttca tcctagtcaa tgtcttatca
     3721 ttgcatacat tttctggatg cttgagccat cagatatcag ctagcttttt aaaatcacct
     3781 ttaagaaagc agtattttct aatagtaaat ttataaacat cttatacatc ttagctttca
     3841 tatataggat tactatgtat tgatttgtaa acattcactg agtttaattt tatttccaca
     3901 gggctcacaa caacatggga gcggaggagg ttccggtgga ggtggttctg gagattacaa
     3961 ggatgacgac gataagggcg attacaagga tgacgacgat aagggagatt acaaggatga
     4021 cgacgataag gtttcaggaa gcggagctac taacttcagc ctgctgaagc aggctggaga
     4081 cgtggaggag aaccctggac ctggatcgtt tcgcatgatt gaacaagatg gattgcacgc
     4141 aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat
     4201 cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt
     4261 caagaccgac ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg
     4321 gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag
     4381 ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc
     4441 tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc
     4501 tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga
     4561 agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga
     4621 actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg
     4681 cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg
     4741 tggccggctg ggtgtggccg accgctatca ggacatagcg ttggctaccc gtgatattgc
     4801 tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc
     4861 cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgat tcgaacatca
     4921 ccaggatgct tatgttctta agtctatatt tgcatataca ttgactcttg atggaaagac
     4981 ttaagaagat caaggtctca ccatttgtcc tcaattcgtg tgaccataag atactgatag
     5041 cattgagtct tgaaatgatt taataatatg agtgaggatt tgctttctcc attagagcat
     5101 taagctaaaa ctatcaacat tttaaaccaa attgccttat ttttcttcca aacttcatat
     5161 atgtctatca ggtaataata ggcttgaaaa ttgatatcct gtggtgctaa agtacagtag
     5221 aaagagagga gaagtgtata catgttttat tttaaattgt acgaaagggg aatttaaaaa
     5281 atatgtaact gctgtttata cattggctcc ttactgctta ttaatctgta ttgtacacat
     5341 gatgaaatga agcagaagct gggagtcggc ctttcctcta gtaaccacca catggctcag
     5401 catctgtgcc aaacataggc gctcctagtc tggtcagtgc caagaggcta ccagaacatg
     5461 gggcaggtgg ctggtgttgg tgtcccagcc taagagccac ctgctgcagt taccatggca
     5521 tgctgagttg atgcaccagg tggcagcagc catccgttat tatttccaat ggagacctag
     5581 cccaggccaa ggtaaagtta gttaatagca ttgggatata gtcactgtaa tggtgctatt
     5641 aacaaacagt caacaccatt gtatttttta acttcgtgtt ctgtatctcc tcagccatgt
     5701 atcttaaata tattttgtcc ttggaaagtc ctctccactg actgtagcct ccaattcact
     5761 ggccgtcgtt cactggccgt cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc
     5821 caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag cgaagaggcc
     5881 cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatgcga tttattcaac
     5941 aaagccgccg tcccgtcaag tcagcgtaat gctctgccag tgttacaacc aattaaccaa
     6001 ttctgattag aaaaactcat cgagcatcaa atgaaactgc aatatattca tatcaggatt
     6061 atcaatacca tatttttgaa aaagccgttt ctgtaatgaa ggagaaaact caccgaggca
     6121 gttccatagg atggcaagat cctggtatcg gtctgcgatt ccgactcgtc caacatcaat
     6181 acaacctatt aatttcccct cgtcaaaaat aaggttatca agtgagaaat caccatgagt
     6241 gacgactgaa tccggtgaga atggcaaaag cttatgcatt tctttccaga cttgttcaac
     6301 aggccagcca ttacgctcgt catcaaaatc actcgcatca accaaaccgt tattcattcg
     6361 tgattgcgcc tgagcgagac gaaatacgcg atcgctgtta aaaggacaat tacaaacagg
     6421 aatcgaatgc aaccggcgca ggaacactgc cagcgcatca acaatatttt cacctgaatc
     6481 aggatattct tctaatacct ggaatgctgt tttcccgggg atcgcagtgg tgagtaacca
     6541 tgcatcatca ggagtacgga taaaatgctt gatggtcgga agaggcataa attccgtcag
     6601 ccagtttagt ctgaccatct catctgtaac atcattggca acgctacctt tgccatgttt
     6661 cagaaacaac tctggcgcat cgggcttccc atacaatcga tagattgtcg cacctgattg
     6721 cccgacatta tcgcgagccc atttataccc atataaatca gcatccatgt tggaatttaa
     6781 tcgcggcttc gagcaagacg tttcccgttg aatatggctc ataacacccc ttgtattact
     6841 gtttatgtaa gcagacagtt ttattgttca tgatgatata tttttatctt gtgcaatgta
     6901 acatcagaga ttttgagaca caacgtggct ttgttgaata aatcgaactt ttgctgagtt
     6961 gaaggatcag atcacgcatc ttcccgacaa cgcagaccgt tccgtggcaa agcaaaagtt
     7021 caaaatcacc aactggtcca cctacaacaa agctctcatc aaccgtggct ccctcacttt
     7081 ctggctggat gatggggcga ttcaggcctg gtatgagtca gcaacacctt cttcacgagg
     7141 cagacctctc gacg
//
