LOCUS       Exported                6906 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Donor vector for 3' FLAG tag of human RFXANK.
ACCESSION   .
VERSION     .
KEYWORDS    pFETCh_RFXANK
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6906)
  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 6906)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6906
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     rep_origin      30..618
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     519..538
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     772..789
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    1089..1110
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        1125..1155
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1163..1179
                     /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     1168..1190
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1187..1203
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1187..1203
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        1380..1582
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1533..1553
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     1579..1603
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             1679..2809
                     /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             3785..3808
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             3812..3835
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             3839..3862
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             3878..3934
                     /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             3947..4741
                     /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(4001..4020)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4611..4630
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5506..5523)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5506..5522)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5515..5537)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             complement(5741..6556)
                     /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     6464..6483
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     6548..6567
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
ORIGIN
        1 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
       61 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
      121 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
      181 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
      241 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
      301 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
      361 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
      421 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
      481 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
      541 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
      601 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
      661 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
      721 accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca
      781 gtgagcgagg aagcggaaga agctcgcaca ttcagcagcg tttttcagcg cgttttcgat
      841 caacgtttca atgttggtat caacaccagg tttaactttg aacttatcgg cactgacggt
      901 tactgatttt gaacttttgc tttgccacgg aacggtctgc gttgtcggga agatgcgtga
      961 tctgatcctt caactcagca aaagttccca atacgcaaac cgcctctccc cgcgcgttgg
     1021 ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc
     1081 aacgcaatta atgtgagtta gctcactcat taggcacccc aggctttaca ctttatgctt
     1141 ccggctcgta tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat
     1201 gaccatgatt acgccaagct tgcatgcctg caggtcgact ctagaggatc cccgggtacc
     1261 gagctcgaat tagcttgcat gcctgcaggt caattccctg gcattatgcc cagtacatga
     1321 ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg
     1381 tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc
     1441 aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt
     1501 tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg
     1561 ggaggtctat ataagcagag ctcgtttagt gaaccgtcag atcgcctgga gacgccatcc
     1621 acgctgtttt gacctccata ggagacaccg ggaccgatcc agcctgggga tcccgaccat
     1681 ggcttcgtac ccctgccatc aacacgcgtc tgcgttcgac caggctgcgc gttctcgcgg
     1741 ccatagcaac cgacgtacgg cgttgcgccc tcgccggcag caagaagcca cggaagtccg
     1801 cctggagcag aaaatgccca cgctactgcg ggtttatata gacggtcctc acgggatggg
     1861 gaaaaccacc accacgcaac tgctggtggc cctgggttcg cgcgacgata tcgtctacgt
     1921 acccgagccg atgacttact ggcaggtgct gggggcttcc gagacaatcg cgaacatcta
     1981 caccacacaa caccgcctcg accagggtga gatatcggcc ggggacgcgg cggtggtaat
     2041 gacaagcgcc cagataacaa tgggcatgcc ttatgccgtg accgacgccg ttctggctcc
     2101 tcatatcggg ggggaggctg ggagctcaca tgccccgccc ccggccctca ccctcatctt
     2161 cgaccgccat cccatcgccg ccctcctgtg ctacccggcc gcgcgatacc ttatgggcag
     2221 catgaccccc caggccgtgc tggcgttcgt ggccctcatc ccgccgacct tgcccggcac
     2281 aaacatcgtg ttgggggccc ttccggagga cagacacatc gaccgcctgg ccaaacgcca
     2341 gcgccccggc gagcggcttg acctggctat gctggccgcg attcgccgcg tttacgggct
     2401 gcttgccaat acggtgcggt atctgcaggg cggcgggtcg tggcgggagg attggggaca
     2461 gctttcgggg acggccgtgc cgccccaggg tgccgagccc cagagcaacg cgggcccacg
     2521 accccatatc ggggacacgt tatttaccct gtttcgggcc cccgagttgc tggcccccaa
     2581 cggcgacctg tacaacgtgt ttgcctgggc cttggacgtc ttggccaaac gcctccgtcc
     2641 catgcacgtc tttatcctgg attacgacca atcgcccgcc ggctgccggg acgccctgct
     2701 gcaacttacc tccgggatgg tccagaccca cgtcaccacc cccggctcca taccgacgat
     2761 ctgcgacctg gcgcgcacgt ttgcccggga gatgggggag gctaactgag gatcccacgt
     2821 cactattgta tactctatat tatactctat gttatactct gtaatcctac tcaataaacg
     2881 tgtcacgcct gtgaaaccgt actaagtctc ccgtgtattc ttatcaccat caggtgacat
     2941 cctcgcccag gctgtcaatc atgccggtat cgattccagt agcaccggcc ccacgctgac
     3001 aacccactct tgcagcgtta gcagcgcccc tcttaacaag ccgaccccca ccagcgtcgc
     3061 ggttactaac actcctctcc ccgacctgca gcccagctgg gattacaggc atgagccacc
     3121 acacccggct gcacctggct aatttttgta gttttagtag agatggggtt tcatcatgtt
     3181 ggtcaggctg gcctttcctg acttcaagtg atccacctac cttggcctcc caaagtgcta
     3241 ggattacagg tgtgagccac caagcccagc ctaattttta aatttttttg gtagagacag
     3301 cgttttgctc tgttgcccaa gctggcctca aattcctgag ctcaagcagt cctccccact
     3361 ggacctccca aagtgctggg attataggtg tgagccacca tgcccaggct catcttttca
     3421 tgaaagtcac aattcacaca ttttgcttgg taggaaatat ttttatatta attctgaaaa
     3481 gctacaaaag tgcattttta caaacttagg ggaagttgag gttcctgggg tgagtcctcg
     3541 gttctcccag ctttgctctg taatgcaggt ctctgcaagg gtccctgttt gtcccctaag
     3601 gggagagcgc aggttggcct gtgcctccac cccactcccg attcaagctc acagcccacc
     3661 ttttccctgc cccatctcag tgcaacaggt gatcgagaac cacatcctca agctcttcca
     3721 gagcaacctg gtgcccgctg accctgaggg gagcggagga ggttccggtg gaggtggttc
     3781 tggagattac aaggatgacg acgataaggg cgattacaag gatgacgacg ataagggaga
     3841 ttacaaggat gacgacgata aggtttcagg aagcggagct actaacttca gcctgctgaa
     3901 gcaggctgga gacgtggagg agaaccctgg acctggatcg tttcgcatga ttgaacaaga
     3961 tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc
     4021 acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc
     4081 ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc
     4141 gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac
     4201 tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc
     4261 tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac
     4321 gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg
     4381 tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct
     4441 cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt
     4501 cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg
     4561 attcatcgac tgtggccggc tgggtgtggc cgaccgctat caggacatag cgttggctac
     4621 ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg
     4681 tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg
     4741 attcgaacat cggggactca gacactcagg gaacaaaatg gtcagccaga gctggggaaa
     4801 cccagaactg acttcaaagg cagcttctgg acaggtggtg ggaggggacc cttcccaaga
     4861 ggaaccaata aaccttctgt gcagaatgag ggactttgct gtgcttcccg gagggcttcc
     4921 tggaggagac agcccctaga ggtctcacac cttctcccca agcacccgca ggtggtagat
     4981 gaccacacgg ccaaaaaagt cagtggcatc ctcaaacgtc accttcagcc ggtccacttc
     5041 agcagctggt atggggaaag tgtgagcgtg ggcagtcaag ggaaactggt gatttctggt
     5101 ttcccacccg caggccccca ccccttccag acccagctca cactccaccg atgtgggcaa
     5161 cctggggaag ctgcaggtcc ctctaagcct ctcacacaga acaggtggtc atcagcacca
     5221 ttttacagaa aaagtggagt cctagaggtc acgtgttcaa gatcacacat cagggttccc
     5281 attcgggttt gggtgagtcc aaagcttcca gaggcacaga ccacccaccc cagaagcagg
     5341 ggcaggatat ctgaagcgag ttgttgtcct cagggtagaa atctacaatc ttgtggagag
     5401 cctgagtgcc ctgtgaacct tcagcagaga aaggcaatga gtgtctgggg tgacttggaa
     5461 agtcctctcc actgactgta gcctccaatt cactggccgt cgttcactgg ccgtcgtttt
     5521 acaacgtcgt gactgggaaa accctggcgt tacccaactt aatcgccttg cagcacatcc
     5581 ccctttcgcc agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt
     5641 gcgcagcctg aatggcgaat gcgatttatt caacaaagcc gccgtcccgt caagtcagcg
     5701 taatgctctg ccagtgttac aaccaattaa ccaattctga ttagaaaaac tcatcgagca
     5761 tcaaatgaaa ctgcaatata ttcatatcag gattatcaat accatatttt tgaaaaagcc
     5821 gtttctgtaa tgaaggagaa aactcaccga ggcagttcca taggatggca agatcctggt
     5881 atcggtctgc gattccgact cgtccaacat caatacaacc tattaatttc ccctcgtcaa
     5941 aaataaggtt atcaagtgag aaatcaccat gagtgacgac tgaatccggt gagaatggca
     6001 aaagcttatg catttctttc cagacttgtt caacaggcca gccattacgc tcgtcatcaa
     6061 aatcactcgc atcaaccaaa ccgttattca ttcgtgattg cgcctgagcg agacgaaata
     6121 cgcgatcgct gttaaaagga caattacaaa caggaatcga atgcaaccgg cgcaggaaca
     6181 ctgccagcgc atcaacaata ttttcacctg aatcaggata ttcttctaat acctggaatg
     6241 ctgttttccc ggggatcgca gtggtgagta accatgcatc atcaggagta cggataaaat
     6301 gcttgatggt cggaagaggc ataaattccg tcagccagtt tagtctgacc atctcatctg
     6361 taacatcatt ggcaacgcta cctttgccat gtttcagaaa caactctggc gcatcgggct
     6421 tcccatacaa tcgatagatt gtcgcacctg attgcccgac attatcgcga gcccatttat
     6481 acccatataa atcagcatcc atgttggaat ttaatcgcgg cttcgagcaa gacgtttccc
     6541 gttgaatatg gctcataaca ccccttgtat tactgtttat gtaagcagag agttttattg
     6601 ttcatgatga tatattttta tcttgtgcaa tgtaacatca gagattttga gacacaacgt
     6661 ggctttgttg aataaatcga acttttgctg agttgaagga tcagatcacg catcttcccg
     6721 acaacgcaga ccgttccgtg gcaaagcaaa agttcaaaat caccaactgg tccacctaca
     6781 acaaagctct catcaaccgt ggctccctca ctttctggct ggatgatggg gcgattcagg
     6841 cctggtatga gtcagcaaca ccttcttcac gaggcagacc tctcgacgga gttccactga
     6901 gcgtca
//
