LOCUS       Exported                6971 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Donor vector for 3' FLAG tag of human KLF9.
ACCESSION   .
VERSION     .
KEYWORDS    pFETCh_KLF9
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6971)
  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 6971)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6971
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     rep_origin      18..606
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     507..526
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     760..777
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    1077..1098
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        1113..1143
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1151..1167
                     /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     1156..1178
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1175..1191
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1175..1191
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        1368..1570
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1521..1541
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     1567..1591
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             1667..2797
                     /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             3830..3853
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             3857..3880
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             3884..3907
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             3923..3979
                     /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             3992..4786
                     /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(4046..4065)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4656..4675
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5559..5576)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5559..5575)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5568..5590)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             complement(5794..6609)
                     /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     6517..6536
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     6601..6620
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
ORIGIN
        1 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
       61 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
      121 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
      181 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
      241 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
      301 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
      361 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
      421 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
      481 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
      541 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
      601 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
      661 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag
      721 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa
      781 gcggaagaag ctcgcacatt cagcagcgtt tttcagcgcg ttttcgatca acgtttcaat
      841 gttggtatca acaccaggtt taactttgaa cttatcggca ctgacggtta ctgattttga
      901 acttttgctt tgccacggaa cggtctgcgt tgtcgggaag atgcgtgatc tgatccttca
      961 actcagcaaa agttcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa
     1021 tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat
     1081 gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg
     1141 ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac
     1201 gccaagcttg catgcctgca ggtcgactct agaggatccc cgggtaccga gctcgaatta
     1261 gcttgcatgc ctgcaggtca attccctggc attatgccca gtacatgacc ttatgggact
     1321 ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt
     1381 ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcaa gtctccaccc
     1441 cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg
     1501 taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat
     1561 aagcagagct cgtttagtga accgtcagat cgcctggaga cgccatccac gctgttttga
     1621 cctccatagg agacaccggg accgatccag cctggggatc ccgaccatgg cttcgtaccc
     1681 ctgccatcaa cacgcgtctg cgttcgacca ggctgcgcgt tctcgcggcc atagcaaccg
     1741 acgtacggcg ttgcgccctc gccggcagca agaagccacg gaagtccgcc tggagcagaa
     1801 aatgcccacg ctactgcggg tttatataga cggtcctcac gggatgggga aaaccaccac
     1861 cacgcaactg ctggtggccc tgggttcgcg cgacgatatc gtctacgtac ccgagccgat
     1921 gacttactgg caggtgctgg gggcttccga gacaatcgcg aacatctaca ccacacaaca
     1981 ccgcctcgac cagggtgaga tatcggccgg ggacgcggcg gtggtaatga caagcgccca
     2041 gataacaatg ggcatgcctt atgccgtgac cgacgccgtt ctggctcctc atatcggggg
     2101 ggaggctggg agctcacatg ccccgccccc ggccctcacc ctcatcttcg accgccatcc
     2161 catcgccgcc ctcctgtgct acccggccgc gcgatacctt atgggcagca tgacccccca
     2221 ggccgtgctg gcgttcgtgg ccctcatccc gccgaccttg cccggcacaa acatcgtgtt
     2281 gggggccctt ccggaggaca gacacatcga ccgcctggcc aaacgccagc gccccggcga
     2341 gcggcttgac ctggctatgc tggccgcgat tcgccgcgtt tacgggctgc ttgccaatac
     2401 ggtgcggtat ctgcagggcg gcgggtcgtg gcgggaggat tggggacagc tttcggggac
     2461 ggccgtgccg ccccagggtg ccgagcccca gagcaacgcg ggcccacgac cccatatcgg
     2521 ggacacgtta tttaccctgt ttcgggcccc cgagttgctg gcccccaacg gcgacctgta
     2581 caacgtgttt gcctgggcct tggacgtctt ggccaaacgc ctccgtccca tgcacgtctt
     2641 tatcctggat tacgaccaat cgcccgccgg ctgccgggac gccctgctgc aacttacctc
     2701 cgggatggtc cagacccacg tcaccacccc cggctccata ccgacgatct gcgacctggc
     2761 gcgcacgttt gcccgggaga tgggggaggc taactgagga tcccacgtca ctattgtata
     2821 ctctatatta tactctatgt tatactctgt aatcctactc aataaacgtg tcacgcctgt
     2881 gaaaccgtac taagtctccc gtgtattctt atcaccatca ggtgacatcc tcgcccaggc
     2941 tgtcaatcat gccggtatcg attccagtag caccggcccc acgctgacaa cccactcttg
     3001 cagcgttagc agcgcccctc ttaacaagcc gacccccacc agcgtcgcgg ttactaacac
     3061 tcctctcccc gacctgcagc ccagctagtt tgggctgcca taacacacaa ccacagactg
     3121 ggtggctgaa acaacagaac tttatttcct cacagttctg gagactgggg tccaaggtag
     3181 aggcagggga cgggttcgtt tcattggagg cctctctcct tggcttgtag atggtggctt
     3241 catccctgtc ttcccaaggt cttccctagt cttcacatgt ctgtgtcctc atctcctctt
     3301 ataaagacac cagtcgtgga ttaaggcctg ccctcatgaa cttgtttaac atcagttgcc
     3361 tctttaaagg ccttaacttc aaatgcagtc attttctgtg gtgctggggg ttaaggcttc
     3421 aacataggga ttttggggag ggggacacat aataatcatc tccgagggat ttatcctttg
     3481 agtcgtaggg actattgacc atgaccgccc ttcggaattt ttttcatgtt ctttgagctg
     3541 tatcctttct gattcctttc tcttaggtga acggcccttt ccctgcacgt ggccagactg
     3601 ccttaaaaag ttctcccgct cagacgagct gacccgccac taccggaccc acactgggga
     3661 aaagcagttc cgctgtccgc tgtgtgagaa gcgcttcatg aggagtgacc acctcacaaa
     3721 gcacgcccgg cggcacaccg agttccaccc cagcatgatc aagcgatcga aaaaggcgct
     3781 ggccaacgca ctagggagcg gaggaggttc cggtggaggt ggttctggag attacaagga
     3841 tgacgacgat aagggcgatt acaaggatga cgacgataag ggagattaca aggatgacga
     3901 cgataaggtt tcaggaagcg gagctactaa cttcagcctg ctgaagcagg ctggagacgt
     3961 ggaggagaac cctggacctg gatcgtttcg catgattgaa caagatggat tgcacgcagg
     4021 ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg
     4081 ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa
     4141 gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct
     4201 ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga
     4261 ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc
     4321 cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac
     4381 ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc
     4441 cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact
     4501 gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga
     4561 tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg
     4621 ccggctgggt gtggccgacc gctatcagga catagcgttg gctacccgtg atattgctga
     4681 agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga
     4741 ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgattcg aacatcacgt
     4801 ggaagccagg gagggatgga ccccgaaagg agaaaagtac tcccaggaaa cagacgcgtg
     4861 aaaactgagc cccagaagag gcacacttga cggcacagga agtcactgct ctttggtcaa
     4921 tattctgatt ttcctctccc tgcattgttt ttaaaaagca cattgtagcc taagatcaaa
     4981 gtcaacaaca ctcggtcccc ttgaagaggc aactctctga acccgtctct gactgttgga
     5041 gggaaggcaa atgcttttgg gttttttggt ttttgttttt gttttttttt ctccttttat
     5101 ttttttgcgg gggagggtag ggagtgggtg ggggggaggg gggtaaggcc aagactgggg
     5161 tagaatttta aagattcaac actggtgtac atatgtccgc tgggtgagtt gacctgtggc
     5221 ctcgcacagt gattctgggc cctttatgct tgctgtctct cagaattgtt ttcttacctt
     5281 ttaatgtaat gacgagtgtg cttcagtttg tttagcaaaa ccactctctt gaatcacgtt
     5341 aacttttgag attaaaaaaa aaaaaacgcc atagcacagc tgtctttatg caagcaagag
     5401 cacatctact ccagcatgat ctgtcatcta aagacttgaa aacaaaaaac agttacttat
     5461 agtcaatggg taagcagagt ctgaatttat actaatcaag acaaaccttg gaaagtcctc
     5521 tccactgact gtagcctcca attcactggc cgtcgttcac tggccgtcgt tttacaacgt
     5581 cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca tccccctttc
     5641 gccagctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc
     5701 ctgaatggcg aatgcgattt attcaacaaa gccgccgtcc cgtcaagtca gcgtaatgct
     5761 ctgccagtgt tacaaccaat taaccaattc tgattagaaa aactcatcga gcatcaaatg
     5821 aaactgcaat atattcatat caggattatc aataccatat ttttgaaaaa gccgtttctg
     5881 taatgaagga gaaaactcac cgaggcagtt ccataggatg gcaagatcct ggtatcggtc
     5941 tgcgattccg actcgtccaa catcaataca acctattaat ttcccctcgt caaaaataag
     6001 gttatcaagt gagaaatcac catgagtgac gactgaatcc ggtgagaatg gcaaaagctt
     6061 atgcatttct ttccagactt gttcaacagg ccagccatta cgctcgtcat caaaatcact
     6121 cgcatcaacc aaaccgttat tcattcgtga ttgcgcctga gcgagacgaa atacgcgatc
     6181 gctgttaaaa ggacaattac aaacaggaat cgaatgcaac cggcgcagga acactgccag
     6241 cgcatcaaca atattttcac ctgaatcagg atattcttct aatacctgga atgctgtttt
     6301 cccggggatc gcagtggtga gtaaccatgc atcatcagga gtacggataa aatgcttgat
     6361 ggtcggaaga ggcataaatt ccgtcagcca gtttagtctg accatctcat ctgtaacatc
     6421 attggcaacg ctacctttgc catgtttcag aaacaactct ggcgcatcgg gcttcccata
     6481 caatcgatag attgtcgcac ctgattgccc gacattatcg cgagcccatt tatacccata
     6541 taaatcagca tccatgttgg aatttaatcg cggcttcgag caagacgttt cccgttgaat
     6601 atggctcata acaccccttg tattactgtt tatgtaagca gacagtttta ttgttcatga
     6661 tgatatattt ttatcttgtg caatgtaaca tcagagattt tgagacacaa cgtggctttg
     6721 ttgaataaat cgaacttttg ctgagttgaa ggatcagatc acgcatcttc ccgacaacgc
     6781 agaccgttcc gtggcaaagc aaaagttcaa aatcaccaac tggtccacct acaacaaagc
     6841 tctcatcaac cgtggctccc tcactttctg gctggatgat ggggcgattc aggcctggta
     6901 tgagtcagca acaccttctt cacgaggcag acctctcgac ggagttccac tgagcgtcag
     6961 accccgtaga a
//
