LOCUS       sequence-112963-        7251 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Homology Arms and 3X Flag with P2A Neo for 3' tagging of human 
            CEBPB.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7251)
  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 7251)
  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_CEBPB
FEATURES             Location/Qualifiers
     source          1..7251
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      54..642
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     543..562
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     796..813
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    1113..1134
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1149..1154,1155..1172,1173..1179)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1187..1203
                     /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     1192..1214
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1211..1227
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1211..1227
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        1404..1606
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1557..1577
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     1603..1627
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             1703..2833
                     /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             3999..4022
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             4026..4049
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             4053..4076
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             4092..4148
                     /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             4161..4955
                     /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(4215..4234)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4825..4844
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5875..5891)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5884..5906)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             complement(6110..6925)
                     /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
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     6833..6852
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     6917..6936
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
ORIGIN
        1 tcgacggagt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
       61 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
      121 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
      181 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
      241 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
      301 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
      361 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
      421 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
      481 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
      541 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
      601 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
      661 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
      721 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc
      781 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagaagctcg cacattcagc
      841 agcgtttttc agcgcgtttt cgatcaacgt ttcaatgttg gtatcaacac caggtttaac
      901 tttgaactta tcggcactga cggttactga ttttgaactt ttgctttgcc acggaacggt
      961 ctgcgttgtc gggaagatgc gtgatctgat ccttcaactc agcaaaagtt cccaatacgc
     1021 aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc
     1081 gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca
     1141 ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa
     1201 caatttcaca caggaaacag ctatgaccat gattacgcca agcttgcatg cctgcaggtc
     1261 gactctagag gatccccggg taccgagctc gaattagctt gcatgcctgc aggtcaattc
     1321 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
     1381 tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat gggcgtggat
     1441 agcggtttga ctcacgggga tttcaagtct ccaccccatt gacgtcaatg ggagtttgtt
     1501 ttggcaccaa aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca
     1561 aatgggcggt aggcgtgtac ggtgggaggt ctatataagc agagctcgtt tagtgaaccg
     1621 tcagatcgcc tggagacgcc atccacgctg ttttgacctc cataggagac accgggaccg
     1681 atccagcctg gggatcccga ccatggcttc gtacccctgc catcaacacg cgtctgcgtt
     1741 cgaccaggct gcgcgttctc gcggccatag caaccgacgt acggcgttgc gccctcgccg
     1801 gcagcaagaa gccacggaag tccgcctgga gcagaaaatg cccacgctac tgcgggttta
     1861 tatagacggt cctcacggga tggggaaaac caccaccacg caactgctgg tggccctggg
     1921 ttcgcgcgac gatatcgtct acgtacccga gccgatgact tactggcagg tgctgggggc
     1981 ttccgagaca atcgcgaaca tctacaccac acaacaccgc ctcgaccagg gtgagatatc
     2041 ggccggggac gcggcggtgg taatgacaag cgcccagata acaatgggca tgccttatgc
     2101 cgtgaccgac gccgttctgg ctcctcatat cgggggggag gctgggagct cacatgcccc
     2161 gcccccggcc ctcaccctca tcttcgaccg ccatcccatc gccgccctcc tgtgctaccc
     2221 ggccgcgcga taccttatgg gcagcatgac cccccaggcc gtgctggcgt tcgtggccct
     2281 catcccgccg accttgcccg gcacaaacat cgtgttgggg gcccttccgg aggacagaca
     2341 catcgaccgc ctggccaaac gccagcgccc cggcgagcgg cttgacctgg ctatgctggc
     2401 cgcgattcgc cgcgtttacg ggctgcttgc caatacggtg cggtatctgc agggcggcgg
     2461 gtcgtggcgg gaggattggg gacagctttc ggggacggcc gtgccgcccc agggtgccga
     2521 gccccagagc aacgcgggcc cacgacccca tatcggggac acgttattta ccctgtttcg
     2581 ggcccccgag ttgctggccc ccaacggcga cctgtacaac gtgtttgcct gggccttgga
     2641 cgtcttggcc aaacgcctcc gtcccatgca cgtctttatc ctggattacg accaatcgcc
     2701 cgccggctgc cgggacgccc tgctgcaact tacctccggg atggtccaga cccacgtcac
     2761 cacccccggc tccataccga cgatctgcga cctggcgcgc acgtttgccc gggagatggg
     2821 ggaggctaac tgaggatccc acgtcactat tgtatactct atattatact ctatgttata
     2881 ctctgtaatc ctactcaata aacgtgtcac gcctgtgaaa ccgtactaag tctcccgtgt
     2941 attcttatca ccatcaggtg acatcctcgc ccaggctgtc aatcatgccg gtatcgattc
     3001 cagtagcacc ggccccacgc tgacaaccca ctcttgcagc gttagcagcg cccctcttaa
     3061 caagccgacc cccaccagcg tcgcggttac taacactcct ctccccgacc tgcagcccag
     3121 cttccccgac ctgcagccca gctatcgact tcagcccgta cctggagccg ctgggcgcgc
     3181 cgcaggcccc ggcgcccgcc acggccacgg acaccttcga ggcggctccg cccgcgcccg
     3241 cccccgcgcc cgcctcctcc gggcagcacc acgacttcct ctccgacctc ttctccgacg
     3301 actacggggg caagaactgc aagaagccgg ccgagtacgg ctacgtgagc ctggggcgcc
     3361 tgggggctgc caagggcgcg ctgcaccccg gctgcttcgc gcccctgcac ccaccgcccc
     3421 cgccgccgcc gccgcccgcc gagctcaagg cggagccggg cttcgagccc gcggactgca
     3481 agcggaagga ggaggccggg gcgccgggcg gcggcgcagg catggcggcg ggcttcccgt
     3541 acgcgctgcg cgcttacctc ggctaccagg cggtgccgag cggcagcagc gggagcctct
     3601 ccacgtcctc ctcgtccagc ccgcccggca cgccgagccc cgctgacgcc aaggcgcccc
     3661 cgaccgcctg ctacgcgggg gccgcgccgg cgccctcgca ggtcaagagc aaggccaaga
     3721 agaccgtgga caagcacagc gacgagtaca agatccggcg cgagcgcaac aacatcgccg
     3781 tgcgcaagag ccgcgacaag gccaagatgc gcaacctgga gacgcagcac aaggtcctgg
     3841 agctcacggc cgagaacgag cggctgcaga agaaggtgga gcagctgtcg cgcgagctca
     3901 gcaccctgcg gaacttgttc aagcagctgc ccgagcccct gctcgcctcc tccggccact
     3961 gcgggagcgg aggaggttcc ggtggaggtg gttctggaga ttacaaggat gacgacgata
     4021 agggcgatta caaggatgac gacgataagg gagattacaa ggatgacgac gataaggttt
     4081 caggaagcgg agctactaac ttcagcctgc tgaagcaggc tggagacgtg gaggagaacc
     4141 ctggacctgg atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg
     4201 cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg
     4261 ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt
     4321 ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg gccacgacgg
     4381 gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat
     4441 tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat
     4501 ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg
     4561 accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg
     4621 atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc
     4681 tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc
     4741 cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg
     4801 tggccgaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg
     4861 gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca
     4921 tcgccttcta tcgccttctt gacgagttct tctgattcga acatcgcgtc cccctgccgg
     4981 ccggggctga gactccgggg agcgcccgcg cccgcgccct cgcccccgcc cccggcggcg
     5041 ccggcaaaac tttggcactg gggcacttgg cagcgcgggg agcccgtcgg taattttaat
     5101 attttattat atatatatat ctatattttt gtccaaacca accgcacatg cagatggggc
     5161 tcccgcccgt ggtgttattt aaagaagaaa cgtctatgtg tacagatgaa tgataaactc
     5221 tctgcttctc cctctgcccc tctccaggcg ccggcgggcg ggccggtttc gaagttgatg
     5281 caatcggttt aaacatggct gaacgcgtgt gtacacggga ctgacgcaac ccacgtgtaa
     5341 ctgtcagccg ggccctgagt aatcgcttaa agatgttcct acgggcttgt tgctgttgat
     5401 gttttgtttt gttttgtttt ttggtctttt tttgtattat aaaaaataat ctatttctat
     5461 gagaaaagag gcgtctgtat attttgggaa tcttttccgt ttcaagcatt aagaacactt
     5521 ttaataaact tttttttgag aatggttaca aagccttttg ggggcagtaa ttggcttttg
     5581 ttttttattt ttttacttta ttttggattt gtaggatttt gtttttgcgt ttctggtgtg
     5641 tagggggttg tgtgtggggg gctgctgtta tttttggagg ttttggtggt tgggtggggg
     5701 tgttgcagct ggtttttctg cctcctctgc tactccccct cccacacaca cagggtctgc
     5761 ttgagatggg gttccagccc cgggggaaag gggagaagag taatgggtca ggcattcagg
     5821 ctcttggaaa gtcctctcca ctgactgtag cctccaattc actggccgtc gttcactggc
     5881 cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt acccaactta atcgccttgc
     5941 agcacatccc cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc
     6001 ccaacagttg cgcagcctga atggcgaatg cgatttattc aacaaagccg ccgtcccgtc
     6061 aagtcagcgt aatgctctgc cagtgttaca accaattaac caattctgat tagaaaaact
     6121 catcgagcat caaatgaaac tgcaatttat tcatatcagg attatcaata ccatattttt
     6181 gaaaaagccg tttctgtaat gaaggagaaa actcaccgag gcagttccat aggatggcaa
     6241 gatcctggta tcggtctgcg attccgactc gtccaacatc aatacaacct attaatttcc
     6301 cctcgtcaaa aataaggtta tcaagtgaga aatcaccatg agtgacgact gaatccggtg
     6361 agaatggcaa aagcttatgc atttctttcc agacttgttc aacaggccag ccattacgct
     6421 cgtcatcaaa atcactcgca tcaaccaaac cgttattcat tcgtgattgc gcctgagcga
     6481 gacgaaatac gcgatcgctg ttaaaaggac aattacaaac aggaatcgaa tgcaaccggc
     6541 gcaggaacac tgccagcgca tcaacaatat tttcacctga atcaggatat tcttctaata
     6601 cctggaatgc tgttttcccg gggatcgcag tggtgagtaa ccatgcatca tcaggagtac
     6661 ggataaaatg cttgatggtc ggaagaggca taaattccgt cagccagttt agtctgacca
     6721 tctcatctgt aacatcattg gcaacgctac ctttgccatg tttcagaaac aactctggcg
     6781 catcgggctt cccatacaat cgatagattg tcgcacctga ttgcccgaca ttatcgcgag
     6841 cccatttata cccatataaa tcagcatcca tgttggaatt taatcgcggc ttcgagcaag
     6901 acgtttcccg ttgaatatgg ctcataacac cccttgtatt actgtttatg taagcagaca
     6961 gttttattgt tcatgatgat atatttttat cttgtgcaat gtaacatcag agattttgag
     7021 acacaacgtg gctttgttga ataaatcgaa cttttgctga gttgaaggat cagatcacgc
     7081 atcttcccga caacgcagac cgttccgtgg caaagcaaaa gttcaaaatc accaactggt
     7141 ccacctacaa caaagctctc atcaaccgtg gctccctcac tttctggctg gatgatgggg
     7201 cgattcaggc ctggtatgag tcagcaacac cttcttcacg aggcagacct c
//
