LOCUS       sequence-171029-        7154 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor vector for 3' FLAG tag of human CEBPG.
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_CEBPG
FEATURES             Location/Qualifiers
     source          1..7154
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      37..625
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     526..545
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     779..796
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    1096..1117
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1132..1137,1138..1155,1156..1162)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1170..1186
                     /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     1175..1197
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1194..1210
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1194..1210
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        1387..1589
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1540..1560
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     1586..1610
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             1686..2816
                     /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             3942..3965
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             3969..3992
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             3996..4019
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             4035..4091
                     /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             4104..4898
                     /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(4158..4177)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4768..4787
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5761..5777)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5770..5792)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             complement(5996..6811)
                     /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     6719..6738
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     6803..6822
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
ORIGIN
        1 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
       61 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
      121 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
      181 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
      241 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
      301 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
      361 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
      421 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
      481 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
      541 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
      601 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
      661 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa
      721 ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag
      781 cgagtcagtg agcgaggaag cggaagaagc tcgcacattc agcagcgttt ttcagcgcgt
      841 tttcgatcaa cgtttcaatg ttggtatcaa caccaggttt aactttgaac ttatcggcac
      901 tgacggttac tgattttgaa cttttgcttt gccacggaac ggtctgcgtt gtcgggaaga
      961 tgcgtgatct gatccttcaa ctcagcaaaa gttcccaata cgcaaaccgc ctctccccgc
     1021 gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag
     1081 tgagcgcaac gcaattaatg tgagttagct cactcattag gcaccccagg ctttacactt
     1141 tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc acacaggaaa
     1201 cagctatgac catgattacg ccaagcttgc atgcctgcag gtcgactcta gaggatcccc
     1261 gggtaccgag ctcgaattag cttgcatgcc tgcaggtcaa ttccctggca ttatgcccag
     1321 tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt
     1381 accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg
     1441 ggatttcaag tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac
     1501 gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg
     1561 tacggtggga ggtctatata agcagagctc gtttagtgaa ccgtcagatc gcctggagac
     1621 gccatccacg ctgttttgac ctccatagga gacaccggga ccgatccagc ctggggatcc
     1681 cgaccatggc ttcgtacccc tgccatcaac acgcgtctgc gttcgaccag gctgcgcgtt
     1741 ctcgcggcca tagcaaccga cgtacggcgt tgcgccctcg ccggcagcaa gaagccacgg
     1801 aagtccgcct ggagcagaaa atgcccacgc tactgcgggt ttatatagac ggtcctcacg
     1861 ggatggggaa aaccaccacc acgcaactgc tggtggccct gggttcgcgc gacgatatcg
     1921 tctacgtacc cgagccgatg acttactggc aggtgctggg ggcttccgag acaatcgcga
     1981 acatctacac cacacaacac cgcctcgacc agggtgagat atcggccggg gacgcggcgg
     2041 tggtaatgac aagcgcccag ataacaatgg gcatgcctta tgccgtgacc gacgccgttc
     2101 tggctcctca tatcgggggg gaggctggga gctcacatgc cccgcccccg gccctcaccc
     2161 tcatcttcga ccgccatccc atcgccgccc tcctgtgcta cccggccgcg cgatacctta
     2221 tgggcagcat gaccccccag gccgtgctgg cgttcgtggc cctcatcccg ccgaccttgc
     2281 ccggcacaaa catcgtgttg ggggcccttc cggaggacag acacatcgac cgcctggcca
     2341 aacgccagcg ccccggcgag cggcttgacc tggctatgct ggccgcgatt cgccgcgttt
     2401 acgggctgct tgccaatacg gtgcggtatc tgcagggcgg cgggtcgtgg cgggaggatt
     2461 ggggacagct ttcggggacg gccgtgccgc cccagggtgc cgagccccag agcaacgcgg
     2521 gcccacgacc ccatatcggg gacacgttat ttaccctgtt tcgggccccc gagttgctgg
     2581 cccccaacgg cgacctgtac aacgtgtttg cctgggcctt ggacgtcttg gccaaacgcc
     2641 tccgtcccat gcacgtcttt atcctggatt acgaccaatc gcccgccggc tgccgggacg
     2701 ccctgctgca acttacctcc gggatggtcc agacccacgt caccaccccc ggctccatac
     2761 cgacgatctg cgacctggcg cgcacgtttg cccgggagat gggggaggct aactgaggat
     2821 cccacgtcac tattgtatac tctatattat actctatgtt atactctgta atcctactca
     2881 ataaacgtgt cacgcctgtg aaaccgtact aagtctcccg tgtattctta tcaccatcag
     2941 gtgacatcct cgcccaggct gtcaatcatg ccggtatcga ttccagtagc accggcccca
     3001 cgctgacaac ccactcttgc agcgttagca gcgcccctct taacaagccg acccccacca
     3061 gcgtcgcggt tactaacact cctctccccg acctgcagcc cagcttttga taacaatcgc
     3121 agaggggcag tgccctcagg gatccaggtg ggctgagttt cgcctgatgg tgcttctgtg
     3181 cttacttacc tttctaagtg aaatgattta gacttcagaa acattggaac agttggcagt
     3241 tgctgtagac ttttctcagc agaatcaatt aatgttggcc tgatgttagg tacgtacaag
     3301 tttgatctca tttgatcctg tcatttcaaa tattttaatg caatttattt tttaactagg
     3361 tacatgtgaa gattttttgg cagcttagcg tggaaaccat tgatcaccct gctctcattt
     3421 ctacctgttc tgtgttggca agggagagtg cccaaatgag caagatatcg cagcaaaaca
     3481 gcactccagg ggtgaacgga attagtgtta tccataccca ggcacatgcc agcggcttac
     3541 agcaggttcc tcagctggtg cctgctggcc ctgggggagg aggcaaagct gtggctccca
     3601 gcaagcagag caaaaagagt tcgcccatgg atcgaaacag tgacgagtat cggcaacgcc
     3661 gagagaggaa caacatggct gtgaaaaaga gccggttgaa aagcaagcag aaagcacaag
     3721 acacactgca gagagtcaat cagctcaaag aagagaatga acggttggaa gcaaaaatca
     3781 aattgctgac caaggaatta agtgtactca aagatttgtt tcttgagcat gcacacaacc
     3841 ttgcagacaa cgtacagtcc attagcactg aaaatacgac agcagatggc gacaatgcag
     3901 gacaggggag cggaggaggt tccggtggag gtggttctgg agattacaag gatgacgacg
     3961 ataagggcga ttacaaggat gacgacgata agggagatta caaggatgac gacgataagg
     4021 tttcaggaag cggagctact aacttcagcc tgctgaagca ggctggagac gtggaggaga
     4081 accctggacc tggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg
     4141 ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg
     4201 atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc
     4261 tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga
     4321 cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc
     4381 tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag
     4441 tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat
     4501 tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg
     4561 tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca
     4621 ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct
     4681 tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg
     4741 gtgtggccga ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg
     4801 gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc
     4861 gcatcgcctt ctatcgcctt cttgacgagt tcttctgatt cgaacatcga gactttagag
     4921 cttgtggctt gaatgttaaa ggtgtgacca ccgacaccac tcatgtcaat ggctgaaagt
     4981 tgtccatttc catgactcaa agacccattg gaggctattt tctgggatca gcactgaaga
     5041 gttgattagc taaaaatgtt agccttgtaa ttcgaatatc tggttttaaa tgatagaggt
     5101 ttttgtggga atcaaaatcc cccaaatgtt aaggtatatg gtaaaaaaag aaatatctgg
     5161 gatcccgatg ttcttaataa atcctgactt cccaagaaat gcttcttttt taagttgaca
     5221 aaaggaatgg ggaactggca ggccgcgcag aaggttcttg gttttaatgg ataggctgaa
     5281 ttggattaag aaaagttgaa tgccacctat ggtaatctat ttgtgatttt cttctaaatt
     5341 atgtattata aattcgtaga gctatagaaa gcaatgagtg tgtaatttgg agtgatttta
     5401 tatatggcat aaactttgtt ttaacataat tagtactgtt ttcccccaaa agtacaagtt
     5461 tttgagtagc aatgtcaggt taagtaaaga aacttcatca catcttatag gtagtgtgtg
     5521 gccaattgac ttaaaaaata caaataacat ttaggaagca aatagattaa acacaaaaat
     5581 aaaactaaag cataggaatt atgtttttga gatacctttg ggcttagatt ggcattgttt
     5641 tattctaaaa acccaactca gtggtgtaga gaaacttgtg taccaaaatt ttagtttctg
     5701 cagatgctct tggaaagtcc tctccactga ctgtagcctc caattcactg gccgtcgttc
     5761 actggccgtc gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg
     5821 ccttgcagca catccccctt tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg
     5881 cccttcccaa cagttgcgca gcctgaatgg cgaatgcgat ttattcaaca aagccgccgt
     5941 cccgtcaagt cagcgtaatg ctctgccagt gttacaacca attaaccaat tctgattaga
     6001 aaaactcatc gagcatcaaa tgaaactgca atatattcat atcaggatta tcaataccat
     6061 atttttgaaa aagccgtttc tgtaatgaag gagaaaactc accgaggcag ttccatagga
     6121 tggcaagatc ctggtatcgg tctgcgattc cgactcgtcc aacatcaata caacctatta
     6181 atttcccctc gtcaaaaata aggttatcaa gtgagaaatc accatgagtg acgactgaat
     6241 ccggtgagaa tggcaaaagc ttatgcattt ctttccagac ttgttcaaca ggccagccat
     6301 tacgctcgtc atcaaaatca ctcgcatcaa ccaaaccgtt attcattcgt gattgcgcct
     6361 gagcgagacg aaatacgcga tcgctgttaa aaggacaatt acaaacagga atcgaatgca
     6421 accggcgcag gaacactgcc agcgcatcaa caatattttc acctgaatca ggatattctt
     6481 ctaatacctg gaatgctgtt ttcccgggga tcgcagtggt gagtaaccat gcatcatcag
     6541 gagtacggat aaaatgcttg atggtcggaa gaggcataaa ttccgtcagc cagtttagtc
     6601 tgaccatctc atctgtaaca tcattggcaa cgctaccttt gccatgtttc agaaacaact
     6661 ctggcgcatc gggcttccca tacaatcgat agattgtcgc acctgattgc ccgacattat
     6721 cgcgagccca tttataccca tataaatcag catccatgtt ggaatttaat cgcggcttcg
     6781 agcaagacgt ttcccgttga atatggctca taacacccct tgtattactg tttatgtaag
     6841 cagacagttt tattgttcat gatgatatat ttttatcttg tgcaatgtaa catcagagat
     6901 tttgagacac aacgtggctt tgttgaataa atcgaacttt tgctgagttg aaggatcaga
     6961 tcacgcatct tcccgacaac gcagaccgtt ccgtggcaaa gcaaaagttc aaaatcacca
     7021 actggtccac ctacaacaaa gctctcatca accgtggctc cctcactttc tggctggatg
     7081 atggggcgat tcaggcctgg tatgagtcag caacaccttc ttcacgaggc agacctctcg
     7141 acggagttcc actg
//
