LOCUS       sequence-171202-        7086 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor vector for 3' FLAG tag of human SP5.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7086)
  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 7086)
  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_SP5
FEATURES             Location/Qualifiers
     source          1..7086
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      50..638
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     539..558
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     792..809
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    1109..1130
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1145..1150,1151..1168,1169..1175)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1183..1199
                     /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     1188..1210
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1207..1223
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1207..1223
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        1400..1602
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1553..1573
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     1599..1623
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             1699..2829
                     /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             3979..4002
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             4006..4029
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             4033..4056
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             4072..4128
                     /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             4141..4935
                     /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(4195..4214)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4805..4824
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5706..5722)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5715..5737)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             complement(5941..6756)
                     /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     6664..6683
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     6748..6767
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
ORIGIN
        1 cggagttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
       61 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
      121 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
      181 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
      241 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
      301 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
      361 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
      421 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
      481 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
      541 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
      601 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
      661 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg
      721 attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa
      781 cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga agctcgcaca ttcagcagcg
      841 tttttcagcg cgttttcgat caacgtttca atgttggtat caacaccagg tttaactttg
      901 aacttatcgg cactgacggt tactgatttt gaacttttgc tttgccacgg aacggtctgc
      961 gttgtcggga agatgcgtga tctgatcctt caactcagca aaagttccca atacgcaaac
     1021 cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact
     1081 ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat taggcacccc
     1141 aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc ggataacaat
     1201 ttcacacagg aaacagctat gaccatgatt acgccaagct tgcatgcctg caggtcgact
     1261 ctagaggatc cccgggtacc gagctcgaat tagcttgcat gcctgcaggt caattccctg
     1321 gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt
     1381 agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg
     1441 gtttgactca cggggatttc aagtctccac cccattgacg tcaatgggag tttgttttgg
     1501 caccaaaatc aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg
     1561 ggcggtaggc gtgtacggtg ggaggtctat ataagcagag ctcgtttagt gaaccgtcag
     1621 atcgcctgga gacgccatcc acgctgtttt gacctccata ggagacaccg ggaccgatcc
     1681 agcctgggga tcccgaccat ggcttcgtac ccctgccatc aacacgcgtc tgcgttcgac
     1741 caggctgcgc gttctcgcgg ccatagcaac cgacgtacgg cgttgcgccc tcgccggcag
     1801 caagaagcca cggaagtccg cctggagcag aaaatgccca cgctactgcg ggtttatata
     1861 gacggtcctc acgggatggg gaaaaccacc accacgcaac tgctggtggc cctgggttcg
     1921 cgcgacgata tcgtctacgt acccgagccg atgacttact ggcaggtgct gggggcttcc
     1981 gagacaatcg cgaacatcta caccacacaa caccgcctcg accagggtga gatatcggcc
     2041 ggggacgcgg cggtggtaat gacaagcgcc cagataacaa tgggcatgcc ttatgccgtg
     2101 accgacgccg ttctggctcc tcatatcggg ggggaggctg ggagctcaca tgccccgccc
     2161 ccggccctca ccctcatctt cgaccgccat cccatcgccg ccctcctgtg ctacccggcc
     2221 gcgcgatacc ttatgggcag catgaccccc caggccgtgc tggcgttcgt ggccctcatc
     2281 ccgccgacct tgcccggcac aaacatcgtg ttgggggccc ttccggagga cagacacatc
     2341 gaccgcctgg ccaaacgcca gcgccccggc gagcggcttg acctggctat gctggccgcg
     2401 attcgccgcg tttacgggct gcttgccaat acggtgcggt atctgcaggg cggcgggtcg
     2461 tggcgggagg attggggaca gctttcgggg acggccgtgc cgccccaggg tgccgagccc
     2521 cagagcaacg cgggcccacg accccatatc ggggacacgt tatttaccct gtttcgggcc
     2581 cccgagttgc tggcccccaa cggcgacctg tacaacgtgt ttgcctgggc cttggacgtc
     2641 ttggccaaac gcctccgtcc catgcacgtc tttatcctgg attacgacca atcgcccgcc
     2701 ggctgccggg acgccctgct gcaacttacc tccgggatgg tccagaccca cgtcaccacc
     2761 cccggctcca taccgacgat ctgcgacctg gcgcgcacgt ttgcccggga gatgggggag
     2821 gctaactgag gatcccacgt cactattgta tactctatat tatactctat gttatactct
     2881 gtaatcctac tcaataaacg tgtcacgcct gtgaaaccgt actaagtctc ccgtgtattc
     2941 ttatcaccat caggtgacat cctcgcccag gctgtcaatc atgccggtat cgattccagt
     3001 agcaccggcc ccacgctgac aacccactct tgcagcgtta gcagcgcccc tcttaacaag
     3061 ccgaccccca ccagcgtcgc ggttactaac actcctctcc ccgacctgca gcccagctag
     3121 ttctcgccgg tcaagatgct gccctcgagc atggcggctc tgcccgccag ctgcgcgccc
     3181 gcctacgtgc cctacgcggc gcaggccgcg ctgccgccag gctactccaa cctgctgcct
     3241 ccgccgccgc caccgccccc gccgcccacc tgccgccagt tgtcacccaa cccggccccc
     3301 gacgacctcc cgtggtggag catcccgcag gcgggcgccg ggccgggggc ctccggggtt
     3361 ccgggaagcg gcctctccgg cgcctgtgcc ggggcccccc acgcgccccg cttccccgcc
     3421 tctgcggccg ctgctgctgc ggccgccgcc gccctacaaa gaggcctggt gttgggcccg
     3481 tcggactttg cgcagtacca gagccagatc gccgcgctgc tgcagaccaa ggcccccctg
     3541 gcggccacgg ccaggaggtg ccgccgctgc cgctgtccca actgccaggc ggcgggcggc
     3601 gcccccgagg cggagccggg gaagaagaag cagcacgtgt gccacgtgcc gggctgcggc
     3661 aaggtgtacg ggaagacgtc gcacctgaag gcgcacctgc gctggcacac gggcgagcga
     3721 cccttcgtgt gcaactggct cttctgcggg aagagcttca cgcgctcgga cgagctgcag
     3781 cggcacctgc ggactcacac gggcgagaag cgctttgcct gtcccgagtg cggcaagcgc
     3841 ttcatgcgca gcgaccacct cgcgaagcac gtcaagactc accagaataa gaagctcaaa
     3901 gtcgctgagg ccggggttaa gcgggaggac gcgcgggacc tggggagcgg aggaggttcc
     3961 ggtggaggtg gttctggaga ttacaaggat gacgacgata agggcgatta caaggatgac
     4021 gacgataagg gagattacaa ggatgacgac gataaggttt caggaagcgg agctactaac
     4081 ttcagcctgc tgaagcaggc tggagacgtg gaggagaacc ctggacctgg atcgtttcgc
     4141 atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc
     4201 ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca
     4261 gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg
     4321 caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg
     4381 ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag
     4441 gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg
     4501 cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc
     4561 atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa
     4621 gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac
     4681 ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat
     4741 ggccgctttt ctggattcat cgactgtggc cggctgggtg tggccgaccg ctatcaggac
     4801 atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc
     4861 ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt
     4921 gacgagttct tctgattcga acatcggagg tggaccccct tcccagcacc tctgcgagag
     4981 atccggggac ctgtgggcag ctggcggagg ggagactcag cagacggacc ctctccgttg
     5041 cctgcctccc aaaatggagc caggcttcca acttccgctg ccttcggaca tagggaccca
     5101 gttcccagga gcggggaggt agggttgggg ctggggcatt tggattgtaa ttgggagctc
     5161 tgccgtacgc cagggcggtt ccaaactcta aaccgttccc accgtcaggg agacctacag
     5221 tttcggggga ccaccctggt ctggccttgt atataggaaa tgctgctgaa ctgaatagaa
     5281 aggaacttgg gagatttgaa acagtgctcg ggttttcgct aggaccggtt tgggctttgt
     5341 acaggttatt taatagcttt gttaaagata attataataa ttataacatt aataaaaatg
     5401 ttgcttttgt cttcagctcc atgcagagct acagcatgat atgtctctgt aaagtgatca
     5461 gcagttgcag cgtgaaaata aatactttaa ctcaggggtc actacaggaa gaccccgttg
     5521 agccggtctc atttgatctt ttcttcttct gggtggctgt actaagcggt cgggtttccc
     5581 cgggaatttg atggttccta tagtctgcct gtgtcggagg cttctttggt ttggaaggat
     5641 ctgggcaaat gtgcttggaa agtcctctcc actgactgta gcctccaatt cactggccgt
     5701 cgttcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt tacccaactt
     5761 aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga ggcccgcacc
     5821 gatcgccctt cccaacagtt gcgcagcctg aatggcgaat gcgatttatt caacaaagcc
     5881 gccgtcccgt caagtcagcg taatgctctg ccagtgttac aaccaattaa ccaattctga
     5941 ttagaaaaac tcatcgagca tcaaatgaaa ctgcaatata ttcatatcag gattatcaat
     6001 accatatttt tgaaaaagcc gtttctgtaa tgaaggagaa aactcaccga ggcagttcca
     6061 taggatggca agatcctggt atcggtctgc gattccgact cgtccaacat caatacaacc
     6121 tattaatttc ccctcgtcaa aaataaggtt atcaagtgag aaatcaccat gagtgacgac
     6181 tgaatccggt gagaatggca aaagcttatg catttctttc cagacttgtt caacaggcca
     6241 gccattacgc tcgtcatcaa aatcactcgc atcaaccaaa ccgttattca ttcgtgattg
     6301 cgcctgagcg agacgaaata cgcgatcgct gttaaaagga caattacaaa caggaatcga
     6361 atgcaaccgg cgcaggaaca ctgccagcgc atcaacaata ttttcacctg aatcaggata
     6421 ttcttctaat acctggaatg ctgttttccc ggggatcgca gtggtgagta accatgcatc
     6481 atcaggagta cggataaaat gcttgatggt cggaagaggc ataaattccg tcagccagtt
     6541 tagtctgacc atctcatctg taacatcatt ggcaacgcta cctttgccat gtttcagaaa
     6601 caactctggc gcatcgggct tcccatacaa tcgatagatt gtcgcacctg attgcccgac
     6661 attatcgcga gcccatttat acccatataa atcagcatcc atgttggaat ttaatcgcgg
     6721 cttcgagcaa gacgtttccc gttgaatatg gctcataaca ccccttgtat tactgtttat
     6781 gtaagcagac agttttattg ttcatgatga tatattttta tcttgtgcaa tgtaacatca
     6841 gagattttga gacacaacgt ggctttgttg aataaatcga acttttgctg agttgaagga
     6901 tcagatcacg catcttcccg acaacgcaga ccgttccgtg gcaaagcaaa agttcaaaat
     6961 caccaactgg tccacctaca acaaagctct catcaaccgt ggctccctca ctttctggct
     7021 ggatgatggg gcgattcagg cctggtatga gtcagcaaca ccttcttcac gaggcagacc
     7081 tctcga
//
