LOCUS       sequence-271679-        7936 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor vector for 3' FLAG tag of human CIC_iso1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7936)
  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 7936)
  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_CIC_iso1
FEATURES             Location/Qualifiers
     source          1..7936
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        121..323
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     274..294
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     320..344
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             420..1550
                     /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             2212..2235
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2239..2262
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2266..2289
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2305..2361
                     /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             2374..3168
                     /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(2428..2447)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3038..3057
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3939..3955)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3948..3970)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             complement(4174..4989)
                     /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     4897..4916
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     4981..5000
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      6707..7295
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7196..7215
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7449..7466
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7766..7787
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7802..7807,7808..7825,7826..7832)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7840..7856
                     /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     7845..7867
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7864..7880
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7864..7880
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 cgagctcgaa ttagcttgca tgcctgcagg tcaattccct ggcattatgc ccagtacatg
       61 accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg
      121 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
      181 caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact
      241 ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt
      301 gggaggtcta tataagcaga gctcgtttag tgaaccgtca gatcgcctgg agacgccatc
      361 cacgctgttt tgacctccat aggagacacc gggaccgatc cagcctgggg atcccgacca
      421 tggcttcgta cccctgccat caacacgcgt ctgcgttcga ccaggctgcg cgttctcgcg
      481 gccatagcaa ccgacgtacg gcgttgcgcc ctcgccggca gcaagaagcc acggaagtcc
      541 gcctggagca gaaaatgccc acgctactgc gggtttatat agacggtcct cacgggatgg
      601 ggaaaaccac caccacgcaa ctgctggtgg ccctgggttc gcgcgacgat atcgtctacg
      661 tacccgagcc gatgacttac tggcaggtgc tgggggcttc cgagacaatc gcgaacatct
      721 acaccacaca acaccgcctc gaccagggtg agatatcggc cggggacgcg gcggtggtaa
      781 tgacaagcgc ccagataaca atgggcatgc cttatgccgt gaccgacgcc gttctggctc
      841 ctcatatcgg gggggaggct gggagctcac atgccccgcc cccggccctc accctcatct
      901 tcgaccgcca tcccatcgcc gccctcctgt gctacccggc cgcgcgatac cttatgggca
      961 gcatgacccc ccaggccgtg ctggcgttcg tggccctcat cccgccgacc ttgcccggca
     1021 caaacatcgt gttgggggcc cttccggagg acagacacat cgaccgcctg gccaaacgcc
     1081 agcgccccgg cgagcggctt gacctggcta tgctggccgc gattcgccgc gtttacgggc
     1141 tgcttgccaa tacggtgcgg tatctgcagg gcggcgggtc gtggcgggag gattggggac
     1201 agctttcggg gacggccgtg ccgccccagg gtgccgagcc ccagagcaac gcgggcccac
     1261 gaccccatat cggggacacg ttatttaccc tgtttcgggc ccccgagttg ctggccccca
     1321 acggcgacct gtacaacgtg tttgcctggg ccttggacgt cttggccaaa cgcctccgtc
     1381 ccatgcacgt ctttatcctg gattacgacc aatcgcccgc cggctgccgg gacgccctgc
     1441 tgcaacttac ctccgggatg gtccagaccc acgtcaccac ccccggctcc ataccgacga
     1501 tctgcgacct ggcgcgcacg tttgcccggg agatggggga ggctaactga ggatcccacg
     1561 tcactattgt atactctata ttatactcta tgttatactc tgtaatccta ctcaataaac
     1621 gtgtcacgcc tgtgaaaccg tactaagtct cccgtgtatt cttatcacca tcaggtgaca
     1681 tcctcgccca ggctgtcaat catgccggta tcgattccag tagcaccggc cccacgctga
     1741 caacccactc ttgcagcgtt agcagcgccc ctcttaacaa gccgaccccc accagcgtcg
     1801 cggttactaa cactcctctc cccgacctgc agcccagctt atgcagacat ctttccctcc
     1861 aaggtttgtc tgcagttgaa gatccgtgag gtgcgccaga agatcatgca ggctgccact
     1921 cccacggagc agccccctgg agctgaggct cctctccctg taccgccccc cactggcacc
     1981 gctgctgccc ctgcccccac tcccagcccc gcagggggcc ctgaccccac ctcacccagc
     2041 tcggactctg gcacggccca ggctgccccg ccactgcctc cacccccaga gtcggggcct
     2101 ggacagcctg gctgggaggg ggctccccag ccctcccccc cacccccagg tccctccaca
     2161 gctgccacag gcagggggag cggaggaggt tccggtggag gtggttctgg agattacaag
     2221 gatgacgacg ataagggcga ttacaaggat gacgacgata agggagatta caaggatgac
     2281 gacgataagg tttcaggaag cggagctact aacttcagcc tgctgaagca ggctggagac
     2341 gtggaggaga accctggacc tggatcgttt cgcatgattg aacaagatgg attgcacgca
     2401 ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc
     2461 ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc
     2521 aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg
     2581 ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg
     2641 gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct
     2701 gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct
     2761 acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa
     2821 gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa
     2881 ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc
     2941 gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt
     3001 ggccggctgg gtgtggccga ccgctatcag gacatagcgt tggctacccg tgatattgct
     3061 gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc
     3121 gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgatt cgaacatcgc
     3181 caggacttat agtaccccct caggacatgg acagtatgtg ggggcaggaa ggttatctcc
     3241 tcccgggtaa agccatttcg tcctctccag tttggggcgg aatgaggcct gctcctcttg
     3301 taaatacccc cttccctcga agctccctcc cggtgctggg gggcagctga ggggctgcag
     3361 gggcagtctc cctcctccaa gcccctgtac ataacctgga gcgtgtgacc ttcagagctt
     3421 ttcactttat gcaaaatggc tcctgtgagg gctgcaagct ggagggtggt gcaggccttg
     3481 ggccacaggg aggcgcctgt ggaatagggg gagttcatgc accccttttt tccccagagg
     3541 ggctggactc aggttagttt gggggtgggg gctcctgcac tttgccacag gcacggggag
     3601 ggttttctcc tcaccccctc tgccctccca acttgggttg tactttctaa gaaggtgatt
     3661 ccccctgccc ttgccccctt ccccagaaca aaacatgttg atcatgtgca atatttctta
     3721 ctgtgccgag aagccgcaat gagcgagatt aaagctgttt aacacacctg tgtggctgta
     3781 ggctcttaac agggagggct gttggttgta ccagtcctgt cctgagttgt catggagcgg
     3841 taggctttcc atgaggtggg ggcaacaatt ccagtaccac tgcccccttg gaaagtcctc
     3901 tccactgact gtagcctcca attcactggc cgtcgttcac tggccgtcgt tttacaacgt
     3961 cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca tccccctttc
     4021 gccagctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc
     4081 ctgaatggcg aatgcgattt attcaacaaa gccgccgtcc cgtcaagtca gcgtaatgct
     4141 ctgccagtgt tacaaccaat taaccaattc tgattagaaa aactcatcga gcatcaaatg
     4201 aaactgcaat atattcatat caggattatc aataccatat ttttgaaaaa gccgtttctg
     4261 taatgaagga gaaaactcac cgaggcagtt ccataggatg gcaagatcct ggtatcggtc
     4321 tgcgattccg actcgtccaa catcaataca acctattaat ttcccctcgt caaaaataag
     4381 gttatcaagt gagaaatcac catgagtgac gactgaatcc ggtgagaatg gcaaaagctt
     4441 atgcatttct ttccagactt gttcaacagg ccagccatta cgctcgtcat caaaatcact
     4501 cgcatcaacc aaaccgttat tcattcgtga ttgcgcctga gcgagacgaa atacgcgatc
     4561 gctgttaaaa ggacaattac aaacaggaat cgaatgcaac cggcgcagga acactgccag
     4621 cgcatcaaca atattttcac ctgaatcagg atattcttct aatacctgga atgctgtttt
     4681 cccggggatc gcagtggtga gtaaccatgc atcatcagga gtacggataa aatgcttgat
     4741 ggtcggaaga ggcataaatt ccgtcagcca gtttagtctg accatctcat ctgtaacatc
     4801 attggcaacg ctacctttgc catgtttcag aaacaactct ggcgcatcgg gcttcccata
     4861 caatcgatag attgtcgcac ctgattgccc gacattatcg cgagcccatt tatacccata
     4921 taaatcagca tccatgttgg aatttaatcg cggcttcgag caagacgttt cccgttgaat
     4981 atggctcata acaccccttg tattactgtt tatgtaagca gacagtttta ttgttcatga
     5041 tgatatattt ttatcttgtg caatgtaaca tcagagattt tgagacacaa cgctgatgaa
     5101 tcccctaatg attttggtaa aaatcattaa gttaaggtgg atacacatct tgtcatatga
     5161 tcaaatggtt tcgcgaaaaa tcaataatca gacaacaaga tgtgcgaact cgatatttta
     5221 cacgactctc tttaccaatt ctgccccgaa ttacacttaa aacgactcaa cagcttaacg
     5281 ttggcttgcc acgcattact tgactgtaaa actctcactc ttaccgaact tggccgtaac
     5341 ctgccaacca aagcgagaac aaaacataac atcaaacgaa tcgaccgatt gttaggtaat
     5401 cgtcacctcc acaaagagcg actcgctgta taccgttggc atgctagctt tatctgttcg
     5461 ggcaatacga tgcccattgt acttgttgac tggtctgata ttcgtgagca aaaacgactt
     5521 atggtattgc gagcttcagt cgcactacac ggtcgttctg ttactcttta tgagaaagcg
     5581 ttcccgcttt cagagcaatg ttcaaagaaa gctcatgacc aatttctagc cgaccttgcg
     5641 agcattctac cgagtaacac cacaccgctc attgtcagtg atgctggctt taaagtgcca
     5701 tggtataaat ccgttgagaa gctgggttgg tactggttaa gtcgagtaag aggaaaagta
     5761 caatatgcag acctaggagc ggaaaactgg aaacctatca gcaacttaca tgatatgtca
     5821 tctagtcact caaagacttt aggctataag aggctgacta aaagcaatcc aatctcatgc
     5881 caaattctat tgtataaatc tcgctctaaa ggccgaaaaa atcagcgctc gacacggact
     5941 cattgtcacc acccgtcacc taaaatctac tcagcgtcgg caaaggagcc atgggttcta
     6001 gcaactaact tacctgttga aattcgaaca cccaaacaac ttgttaatat ctattcgaag
     6061 cgaatgcaga ttgaagaaac cttccgagac ttgaaaagtc ctgcctacgg actaggccta
     6121 cgccatagcc gaacgagcag ctcagagcgt tttgatatca tgctgctaat cgccctgatg
     6181 cttcaactaa catgttggct tgcgggcgtt catgctcaga aacaaggttg ggacaagcac
     6241 ttccaggcta acacagtcag aaatcgaaac gtactctcaa cagttcgctt aggcatggaa
     6301 gttttgcggc attctggcta cacaataaca agggaagact tactcgtggc tgcaacccta
     6361 ctagctcaaa atttattcac acatggttac gctttgggga aattatgagg ggatctctca
     6421 ggacacaacg tggctttgtt gaataaatcg aacttttgct gagttgaagg atcagatcac
     6481 gcatcttccc gacaacgcag accgttccgt ggcaaagcaa aagttcaaaa tcaccaactg
     6541 gtccacctac aacaaagctc tcatcaaccg tggctccctc actttctggc tggatgatgg
     6601 ggcgattcag gcctggtatg agtcagcaac accttcttca cgaggcagac ctctcgacgg
     6661 agttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     6721 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     6781 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     6841 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     6901 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     6961 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     7021 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     7081 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     7141 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     7201 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     7261 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     7321 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
     7381 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga
     7441 ccgagcgcag cgagtcagtg agcgaggaag cggaagaagc tcgcacattc agcagcgttt
     7501 ttcagcgcgt tttcgatcaa cgtttcaatg ttggtatcaa caccaggttt aactttgaac
     7561 ttatcggcac tgacggttac tgattttgaa cttttgcttt gccacggaac ggtctgcgtt
     7621 gtcgggaaga tgcgtgatct gatccttcaa ctcagcaaaa gttcccaata cgcaaaccgc
     7681 ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga
     7741 aagcgggcag tgagcgcaac gcaattaatg tgagttagct cactcattag gcaccccagg
     7801 ctttacactt tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc
     7861 acacaggaaa cagctatgac catgattacg ccaagcttgc atgcctgcag gtcgactcta
     7921 gaggatcccc gggtac
//
