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             354..377
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             381..404
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             408..431
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             447..503
                     /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             516..1310
                     /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(570..589)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1180..1199
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(2081..2098)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2081..2097)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2090..2112)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             complement(2316..3131)
                     /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     3039..3058
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3123..3142
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      3511..4099
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4000..4019
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4253..4270
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4570..4591
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4606..4636
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4644..4660
                     /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     4649..4671
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4668..4684
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4668..4684
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        4861..5063
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5014..5034
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     5060..5084
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             5160..6290
                     /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 tcaaggggct gggcgccgcg caccccgacc tccgcgcgag tggcggcagc ggcgcgggca
       61 aggccaagaa gtcggtggac aagaacagca acgagtaccg ggtgcggcgc gagcgcaaca
      121 acatcgcggt gcgcaagagc cgcgacaagg ccaagcagcg caacgtggag acgcagcaga
      181 aggtgctgga gctgaccagt gacaatgacc gcctgcgcaa gcgggtggaa cagctgagcc
      241 gcgaactgga cacgctgcgg ggcatcttcc gccagctgcc agagagctcc ttggtcaagg
      301 ccatgggcaa ctgcgcaggg agcggaggag gttccggtgg aggtggttct ggagattaca
      361 aggatgacga cgataagggc gattacaagg atgacgacga taagggagat tacaaggatg
      421 acgacgataa ggtttcagga agcggagcta ctaacttcag cctgctgaag caggctggag
      481 acgtggagga gaaccctgga cctggatcgt ttcgcatgat tgaacaagat ggattgcacg
      541 caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa
      601 tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg
      661 tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg cggctatcgt
      721 ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa
      781 gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc
      841 ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg
      901 ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg
      961 aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg
     1021 aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg
     1081 gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact
     1141 gtggccggct gggtgtggcc gaccgctatc aggacatagc gttggctacc cgtgatattg
     1201 ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc
     1261 ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga ttcgaacatc
     1321 gcgcggctgt gggaccgccc tgggccagcc tccggcgggg acccagggag tggtttgggg
     1381 tcgccggatc tcgaggcttg cccgagccgt gcgagccagg actaggagat tccggtgcct
     1441 cctgaaagcc tggcctgctc cgcgtgtccc ctcccttcct ctgcgccgga cttggtgcgt
     1501 ctaagatgag ggggccaggc ggtggcttct ccctgcgagg aggggagaat tcttggggct
     1561 gagctgggag cccggcaact ctagtattta ggataacctt gtgccttgga aatgcaaact
     1621 caccgctcca atgcctactg agtaggggga gcaaatcgtg ccttgtcatt ttatttggag
     1681 gtttcctgcc tccttcccga ggctacagca gacccccatg agagaaggag gggagcaggc
     1741 ccgtggcagg aggagggctc agggagctga gatcccgaca agcccgccag ccccagccgc
     1801 tcctccacgc ctgtccttag aaaggggtgg aaacataggg acttggggct tggaacctaa
     1861 ggttgttccc ctagttctac atgaaggtgg agggtctcta gttccacgcc tctcccacct
     1921 ccctccgcac acaccccacc ccagcctgct ataggctggg cttccccttg gggcggaact
     1981 cactgcgatg ggggtcacca ggtgaccagt gggagccccc accccgagct tggaaagtcc
     2041 tctccactga ctgtagcctc caattcactg gccgtcgttc actggccgtc gttttacaac
     2101 gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca catccccctt
     2161 tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca
     2221 gcctgaatgg cgaatgcgat ttattcaaca aagccgccgt cccgtcaagt cagcgtaatg
     2281 ctctgccagt gttacaacca attaaccaat tctgattaga aaaactcatc gagcatcaaa
     2341 tgaaactgca atttattcat atcaggatta tcaataccat atttttgaaa aagccgtttc
     2401 tgtaatgaag gagaaaactc accgaggcag ttccatagga tggcaagatc ctggtatcgg
     2461 tctgcgattc cgactcgtcc aacatcaata caacctatta atttcccctc gtcaaaaata
     2521 aggttatcaa gtgagaaatc accatgagtg acgactgaat ccggtgagaa tggcaaaagc
     2581 ttatgcattt ctttccagac ttgttcaaca ggccagccat tacgctcgtc atcaaaatca
     2641 ctcgcatcaa ccaaaccgtt attcattcgt gattgcgcct gagcgagacg aaatacgcga
     2701 tcgctgttaa aaggacaatt acaaacagga atcgaatgca accggcgcag gaacactgcc
     2761 agcgcatcaa caatattttc acctgaatca ggatattctt ctaatacctg gaatgctgtt
     2821 ttcccgggga tcgcagtggt gagtaaccat gcatcatcag gagtacggat aaaatgcttg
     2881 atggtcggaa gaggcataaa ttccgtcagc cagtttagtc tgaccatctc atctgtaaca
     2941 tcattggcaa cgctaccttt gccatgtttc agaaacaact ctggcgcatc gggcttccca
     3001 tacaatcgat agattgtcgc acctgattgc ccgacattat cgcgagccca tttataccca
     3061 tataaatcag catccatgtt ggaatttaat cgcggcttcg agcaagacgt ttcccgttga
     3121 atatggctca taacacccct tgtattactg tttatgtaag cagacagttt tattgttcat
     3181 gatgatatat ttttatcttg tgcaatgtaa catcagagat tttgagacac aacgtggctt
     3241 tgttgaataa atcgaacttt tgctgagttg aaggatcaga tcacgcatct tcccgacaac
     3301 gcagaccgtt ccgtggcaaa gcaaaagttc aaaatcacca actggtccac ctacaacaaa
     3361 gctctcatca accgtggctc cctcactttc tggctggatg atggggcgat tcaggcctgg
     3421 tatgagtcag caacaccttc ttcacgaggc agacctctcg acggagttcc actgagcgtc
     3481 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     3541 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     3601 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     3661 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     3721 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     3781 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     3841 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     3901 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     3961 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     4021 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     4081 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     4141 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
     4201 taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc
     4261 agtgagcgag gaagcggaag aagctcgcac attcagcagc gtttttcagc gcgttttcga
     4321 tcaacgtttc aatgttggta tcaacaccag gtttaacttt gaacttatcg gcactgacgg
     4381 ttactgattt tgaacttttg ctttgccacg gaacggtctg cgttgtcggg aagatgcgtg
     4441 atctgatcct tcaactcagc aaaagttccc aatacgcaaa ccgcctctcc ccgcgcgttg
     4501 gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg
     4561 caacgcaatt aatgtgagtt agctcactca ttaggcaccc caggctttac actttatgct
     4621 tccggctcgt atgttgtgtg gaattgtgag cggataacaa tttcacacag gaaacagcta
     4681 tgaccatgat tacgccaagc ttgcatgcct gcaggtcgac tctagaggat ccccgggtac
     4741 cgagctcgaa ttagcttgca tgcctgcagg tcaattccct ggcattatgc ccagtacatg
     4801 accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg
     4861 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
     4921 caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact
     4981 ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt
     5041 gggaggtcta tataagcaga gctcgtttag tgaaccgtca gatcgcctgg agacgccatc
     5101 cacgctgttt tgacctccat aggagacacc gggaccgatc cagcctgggg atcccgacca
     5161 tggcttcgta cccctgccat caacacgcgt ctgcgttcga ccaggctgcg cgttctcgcg
     5221 gccatagcaa ccgacgtacg gcgttgcgcc ctcgccggca gcaagaagcc acggaagtcc
     5281 gcctggagca gaaaatgccc acgctactgc gggtttatat agacggtcct cacgggatgg
     5341 ggaaaaccac caccacgcaa ctgctggtgg ccctgggttc gcgcgacgat atcgtctacg
     5401 tacccgagcc gatgacttac tggcaggtgc tgggggcttc cgagacaatc gcgaacatct
     5461 acaccacaca acaccgcctc gaccagggtg agatatcggc cggggacgcg gcggtggtaa
     5521 tgacaagcgc ccagataaca atgggcatgc cttatgccgt gaccgacgcc gttctggctc
     5581 ctcatatcgg gggggaggct gggagctcac atgccccgcc cccggccctc accctcatct
     5641 tcgaccgcca tcccatcgcc gccctcctgt gctacccggc cgcgcgatac cttatgggca
     5701 gcatgacccc ccaggccgtg ctggcgttcg tggccctcat cccgccgacc ttgcccggca
     5761 caaacatcgt gttgggggcc cttccggagg acagacacat cgaccgcctg gccaaacgcc
     5821 agcgccccgg cgagcggctt gacctggcta tgctggccgc gattcgccgc gtttacgggc
     5881 tgcttgccaa tacggtgcgg tatctgcagg gcggcgggtc gtggcgggag gattggggac
     5941 agctttcggg gacggccgtg ccgccccagg gtgccgagcc ccagagcaac gcgggcccac
     6001 gaccccatat cggggacacg ttatttaccc tgtttcgggc ccccgagttg ctggccccca
     6061 acggcgacct gtacaacgtg tttgcctggg ccttggacgt cttggccaaa cgcctccgtc
     6121 ccatgcacgt ctttatcctg gattacgacc aatcgcccgc cggctgccgg gacgccctgc
     6181 tgcaacttac ctccgggatg gtccagaccc acgtcaccac ccccggctcc ataccgacga
     6241 tctgcgacct ggcgcgcacg tttgcccggg agatggggga ggctaactga ggatcccacg
     6301 tcactattgt atactctata ttatactcta tgttatactc tgtaatccta ctcaataaac
     6361 gtgtcacgcc tgtgaaaccg tactaagtct cccgtgtatt cttatcacca tcaggtgaca
     6421 tcctcgccca ggctgtcaat catgccggta tcgattccag tagcaccggc cccacgctga
     6481 caacccactc ttgcagcgtt agcagcgccc ctcttaacaa gccgaccccc accagcgtcg
     6541 cggttactaa cactcctctc cccgacctgc agcccagct
//
