LOCUS       Exported                6579 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Donor vector for 3' FLAG tag of human CEBPA.
ACCESSION   .
VERSION     .
KEYWORDS    pFETCh_CEBPA
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6579)
  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 6579)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6579
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     CDS             371..394
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             398..421
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             425..448
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             464..520
                     /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             533..1327
                     /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(587..606)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1197..1216
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(2098..2115)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2098..2114)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2107..2129)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             complement(2333..3148)
                     /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     3056..3075
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3140..3159
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      3528..4116
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4017..4036
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4270..4287
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4587..4608
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4623..4653
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4661..4677
                     /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     4666..4688
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4685..4701
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4685..4701
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        4878..5080
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5031..5051
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     5077..5101
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             5177..6307
                     /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"
ORIGIN
        1 cgacctgcag cccagcttca aggggctggg cgccgcgcac cccgacctcc gcgcgagtgg
       61 cggcagcggc gcgggcaagg ccaagaagtc ggtggacaag aacagcaacg agtaccgggt
      121 gcggcgcgag cgcaacaaca tcgcggtgcg caagagccgc gacaaggcca agcagcgcaa
      181 cgtggagacg cagcagaagg tgctggagct gaccagtgac aatgaccgcc tgcgcaagcg
      241 ggtggaacag ctgagccgcg aactggacac gctgcggggc atcttccgcc agctgccaga
      301 gagctccttg gtcaaggcca tgggcaactg cgcagggagc ggaggaggtt ccggtggagg
      361 tggttctgga gattacaagg atgacgacga taagggcgat tacaaggatg acgacgataa
      421 gggagattac aaggatgacg acgataaggt ttcaggaagc ggagctacta acttcagcct
      481 gctgaagcag gctggagacg tggaggagaa ccctggacct ggatcgtttc gcatgattga
      541 acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga
      601 ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg
      661 gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga
      721 ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt
      781 tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct
      841 gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct
      901 gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg
      961 agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca
     1021 ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga
     1081 tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt
     1141 ttctggattc atcgactgtg gccggctggg tgtggccgac cgctatcagg acatagcgtt
     1201 ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct
     1261 ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt
     1321 cttctgattc gaacatcgcg cggctgtggg accgccctgg gccagcctcc ggcggggacc
     1381 cagggagtgg tttggggtcg ccggatctcg aggcttgccc gagccgtgcg agccaggact
     1441 aggagattcc ggtgcctcct gaaagcctgg cctgctccgc gtgtcccctc ccttcctctg
     1501 cgccggactt ggtgcgtcta agatgagggg gccaggcggt ggcttctccc tgcgaggagg
     1561 ggagaattct tggggctgag ctgggagccc ggcaactcta gtatttagga taaccttgtg
     1621 ccttggaaat gcaaactcac cgctccaatg cctactgagt agggggagca aatcgtgcct
     1681 tgtcatttta tttggaggtt tcctgcctcc ttcccgaggc tacagcagac ccccatgaga
     1741 gaaggagggg agcaggcccg tggcaggagg agggctcagg gagctgagat cccgacaagc
     1801 ccgccagccc cagccgctcc tccacgcctg tccttagaaa ggggtggaaa catagggact
     1861 tggggcttgg aacctaaggt tgttccccta gttctacatg aaggtggagg gtctctagtt
     1921 ccacgcctct cccacctccc tccgcacaca ccccacccca gcctgctata ggctgggctt
     1981 ccccttgggg cggaactcac tgcgatgggg gtcaccaggt gaccagtggg agcccccacc
     2041 ccgagcttgg aaagtcctct ccactgactg tagcctccaa ttcactggcc gtcgttcact
     2101 ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc gttacccaac ttaatcgcct
     2161 tgcagcacat ccccctttcg ccagctggcg taatagcgaa gaggcccgca ccgatcgccc
     2221 ttcccaacag ttgcgcagcc tgaatggcga atgcgattta ttcaacaaag ccgccgtccc
     2281 gtcaagtcag cgtaatgctc tgccagtgtt acaaccaatt aaccaattct gattagaaaa
     2341 actcatcgag catcaaatga aactgcaata tattcatatc aggattatca ataccatatt
     2401 tttgaaaaag ccgtttctgt aatgaaggag aaaactcacc gaggcagttc cataggatgg
     2461 caagatcctg gtatcggtct gcgattccga ctcgtccaac atcaatacaa cctattaatt
     2521 tcccctcgtc aaaaataagg ttatcaagtg agaaatcacc atgagtgacg actgaatccg
     2581 gtgagaatgg caaaagctta tgcatttctt tccagacttg ttcaacaggc cagccattac
     2641 gctcgtcatc aaaatcactc gcatcaacca aaccgttatt cattcgtgat tgcgcctgag
     2701 cgagacgaaa tacgcgatcg ctgttaaaag gacaattaca aacaggaatc gaatgcaacc
     2761 ggcgcaggaa cactgccagc gcatcaacaa tattttcacc tgaatcagga tattcttcta
     2821 atacctggaa tgctgttttc ccggggatcg cagtggtgag taaccatgca tcatcaggag
     2881 tacggataaa atgcttgatg gtcggaagag gcataaattc cgtcagccag tttagtctga
     2941 ccatctcatc tgtaacatca ttggcaacgc tacctttgcc atgtttcaga aacaactctg
     3001 gcgcatcggg cttcccatac aatcgataga ttgtcgcacc tgattgcccg acattatcgc
     3061 gagcccattt atacccatat aaatcagcat ccatgttgga atttaatcgc ggcttcgagc
     3121 aagacgtttc ccgttgaata tggctcataa caccccttgt attactgttt atgtaagcag
     3181 acagttttat tgttcatgat gatatatttt tatcttgtgc aatgtaacat cagagatttt
     3241 gagacacaac gtggctttgt tgaataaatc gaacttttgc tgagttgaag gatcagatca
     3301 cgcatcttcc cgacaacgca gaccgttccg tggcaaagca aaagttcaaa atcaccaact
     3361 ggtccaccta caacaaagct ctcatcaacc gtggctccct cactttctgg ctggatgatg
     3421 gggcgattca ggcctggtat gagtcagcaa caccttcttc acgaggcaga cctctcgacg
     3481 gagttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     3541 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     3601 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     3661 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     3721 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     3781 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     3841 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     3901 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     3961 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     4021 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     4081 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     4141 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     4201 tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg
     4261 accgagcgca gcgagtcagt gagcgaggaa gcggaagaag ctcgcacatt cagcagcgtt
     4321 tttcagcgcg ttttcgatca acgtttcaat gttggtatca acaccaggtt taactttgaa
     4381 cttatcggca ctgacggtta ctgattttga acttttgctt tgccacggaa cggtctgcgt
     4441 tgtcgggaag atgcgtgatc tgatccttca actcagcaaa agttcccaat acgcaaaccg
     4501 cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg
     4561 aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta ggcaccccag
     4621 gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt
     4681 cacacaggaa acagctatga ccatgattac gccaagcttg catgcctgca ggtcgactct
     4741 agaggatccc cgggtaccga gctcgaatta gcttgcatgc ctgcaggtca attccctggc
     4801 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
     4861 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
     4921 ttgactcacg gggatttcaa gtctccaccc cattgacgtc aatgggagtt tgttttggca
     4981 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg
     5041 cggtaggcgt gtacggtggg aggtctatat aagcagagct cgtttagtga accgtcagat
     5101 cgcctggaga cgccatccac gctgttttga cctccatagg agacaccggg accgatccag
     5161 cctggggatc ccgaccatgg cttcgtaccc ctgccatcaa cacgcgtctg cgttcgacca
     5221 ggctgcgcgt tctcgcggcc atagcaaccg acgtacggcg ttgcgccctc gccggcagca
     5281 agaagccacg gaagtccgcc tggagcagaa aatgcccacg ctactgcggg tttatataga
     5341 cggtcctcac gggatgggga aaaccaccac cacgcaactg ctggtggccc tgggttcgcg
     5401 cgacgatatc gtctacgtac ccgagccgat gacttactgg caggtgctgg gggcttccga
     5461 gacaatcgcg aacatctaca ccacacaaca ccgcctcgac cagggtgaga tatcggccgg
     5521 ggacgcggcg gtggtaatga caagcgccca gataacaatg ggcatgcctt atgccgtgac
     5581 cgacgccgtt ctggctcctc atatcggggg ggaggctggg agctcacatg ccccgccccc
     5641 ggccctcacc ctcatcttcg accgccatcc catcgccgcc ctcctgtgct acccggccgc
     5701 gcgatacctt atgggcagca tgacccccca ggccgtgctg gcgttcgtgg ccctcatccc
     5761 gccgaccttg cccggcacaa acatcgtgtt gggggccctt ccggaggaca gacacatcga
     5821 ccgcctggcc aaacgccagc gccccggcga gcggcttgac ctggctatgc tggccgcgat
     5881 tcgccgcgtt tacgggctgc ttgccaatac ggtgcggtat ctgcagggcg gcgggtcgtg
     5941 gcgggaggat tggggacagc tttcggggac ggccgtgccg ccccagggtg ccgagcccca
     6001 gagcaacgcg ggcccacgac cccatatcgg ggacacgtta tttaccctgt ttcgggcccc
     6061 cgagttgctg gcccccaacg gcgacctgta caacgtgttt gcctgggcct tggacgtctt
     6121 ggccaaacgc ctccgtccca tgcacgtctt tatcctggat tacgaccaat cgcccgccgg
     6181 ctgccgggac gccctgctgc aacttacctc cgggatggtc cagacccacg tcaccacccc
     6241 cggctccata ccgacgatct gcgacctggc gcgcacgttt gcccgggaga tgggggaggc
     6301 taactgagga tcccacgtca ctattgtata ctctatatta tactctatgt tatactctgt
     6361 aatcctactc aataaacgtg tcacgcctgt gaaaccgtac taagtctccc gtgtattctt
     6421 atcaccatca ggtgacatcc tcgcccaggc tgtcaatcat gccggtatcg attccagtag
     6481 caccggcccc acgctgacaa cccactcttg cagcgttagc agcgcccctc ttaacaagcc
     6541 gacccccacc agcgtcgcgg ttactaacac tcctctccc
//
