LOCUS       Exported                7003 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Donor vector for 3' FLAG tag of human KDM6A.
ACCESSION   .
VERSION     .
KEYWORDS    pFETCh_KDM6A
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7003)
  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 7003)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7003
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     rep_origin      82..670
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     571..590
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     824..841
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    1141..1162
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        1177..1207
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1215..1231
                     /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     1220..1242
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1239..1255
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1239..1255
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        1432..1634
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1585..1605
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     1631..1655
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             1731..2861
                     /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             3928..3951
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             3955..3978
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             3982..4005
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             4021..4077
                     /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             4090..4884
                     /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(4144..4163)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4754..4773
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5655..5672)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5655..5671)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5664..5686)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             complement(5890..6705)
                     /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     6613..6632
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     6697..6716
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
ORIGIN
        1 gcaacacctt cttcacgagg cagacctctc gacggagttc cactgagcgt cagaccccgt
       61 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
      121 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
      181 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
      241 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
      301 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
      361 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
      421 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
      481 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
      541 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
      601 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
      661 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
      721 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
      781 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
      841 ggaagcggaa gaagctcgca cattcagcag cgtttttcag cgcgttttcg atcaacgttt
      901 caatgttggt atcaacacca ggtttaactt tgaacttatc ggcactgacg gttactgatt
      961 ttgaactttt gctttgccac ggaacggtct gcgttgtcgg gaagatgcgt gatctgatcc
     1021 ttcaactcag caaaagttcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca
     1081 ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat
     1141 taatgtgagt tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg
     1201 tatgttgtgt ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga
     1261 ttacgccaag cttgcatgcc tgcaggtcga ctctagagga tccccgggta ccgagctcga
     1321 attagcttgc atgcctgcag gtcaattccc tggcattatg cccagtacat gaccttatgg
     1381 gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg
     1441 ttttggcagt acatcaatgg gcgtggatag cggtttgact cacggggatt tcaagtctcc
     1501 accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat
     1561 gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct
     1621 atataagcag agctcgttta gtgaaccgtc agatcgcctg gagacgccat ccacgctgtt
     1681 ttgacctcca taggagacac cgggaccgat ccagcctggg gatcccgacc atggcttcgt
     1741 acccctgcca tcaacacgcg tctgcgttcg accaggctgc gcgttctcgc ggccatagca
     1801 accgacgtac ggcgttgcgc cctcgccggc agcaagaagc cacggaagtc cgcctggagc
     1861 agaaaatgcc cacgctactg cgggtttata tagacggtcc tcacgggatg gggaaaacca
     1921 ccaccacgca actgctggtg gccctgggtt cgcgcgacga tatcgtctac gtacccgagc
     1981 cgatgactta ctggcaggtg ctgggggctt ccgagacaat cgcgaacatc tacaccacac
     2041 aacaccgcct cgaccagggt gagatatcgg ccggggacgc ggcggtggta atgacaagcg
     2101 cccagataac aatgggcatg ccttatgccg tgaccgacgc cgttctggct cctcatatcg
     2161 ggggggaggc tgggagctca catgccccgc ccccggccct caccctcatc ttcgaccgcc
     2221 atcccatcgc cgccctcctg tgctacccgg ccgcgcgata ccttatgggc agcatgaccc
     2281 cccaggccgt gctggcgttc gtggccctca tcccgccgac cttgcccggc acaaacatcg
     2341 tgttgggggc ccttccggag gacagacaca tcgaccgcct ggccaaacgc cagcgccccg
     2401 gcgagcggct tgacctggct atgctggccg cgattcgccg cgtttacggg ctgcttgcca
     2461 atacggtgcg gtatctgcag ggcggcgggt cgtggcggga ggattgggga cagctttcgg
     2521 ggacggccgt gccgccccag ggtgccgagc cccagagcaa cgcgggccca cgaccccata
     2581 tcggggacac gttatttacc ctgtttcggg cccccgagtt gctggccccc aacggcgacc
     2641 tgtacaacgt gtttgcctgg gccttggacg tcttggccaa acgcctccgt cccatgcacg
     2701 tctttatcct ggattacgac caatcgcccg ccggctgccg ggacgccctg ctgcaactta
     2761 cctccgggat ggtccagacc cacgtcacca cccccggctc cataccgacg atctgcgacc
     2821 tggcgcgcac gtttgcccgg gagatggggg aggctaactg aggatcccac gtcactattg
     2881 tatactctat attatactct atgttatact ctgtaatcct actcaataaa cgtgtcacgc
     2941 ctgtgaaacc gtactaagtc tcccgtgtat tcttatcacc atcaggtgac atcctcgccc
     3001 aggctgtcaa tcatgccggt atcgattcca gtagcaccgg ccccacgctg acaacccact
     3061 cttgcagcgt tagcagcgcc cctcttaaca agccgacccc caccagcgtc gcggttacta
     3121 acactcctct ccccgacctg cagcccagct tgtccttgcc agaagtatta acctcatggg
     3181 agtatttaat ggagcttaac ttcctggtat attatgtaac cttatgaaat tcaagtatac
     3241 ttataaattg cacagttttt aatttcagac agcagatgct tgggggaagc attttacttt
     3301 cgcagatcaa tagctttatg aatagaaaca cagagataat ttatgaattg ttgattttct
     3361 ccagtacttg tctcccacac atatgtgatc agaaactaac tttttgaaat ttactttctg
     3421 atattttcac ttgaatggtc ttcaagattg cttgaagaag aaaattatct gaaatatgtt
     3481 gaagaaaata tattacctag ctgagataga ttctgaaggc agtcactagt gaaattgtaa
     3541 tttgttaaat tattttgcaa gaagccccct ttttttaagc aatcttaaaa aatggtaaac
     3601 gtaataacat tccaagagtt tttgactaat tggtgaagaa gtatagaaaa ttgcaagtac
     3661 actgagtagg ataatggtct tagattctta ggaagattgg ctgaatggga gatagtaagg
     3721 taggctgtac ttttatgttg catagcaggc tgttgaactt tttgagagtt atttccccta
     3781 acttcacaag cagactatat gtttgtagcc atgagctatt aacaatttgt atatcaaaat
     3841 aacctacctt acttttattt tcaggctcct ccattaccat ccgcctcatc tgggagcgga
     3901 ggaggttccg gtggaggtgg ttctggagat tacaaggatg acgacgataa gggcgattac
     3961 aaggatgacg acgataaggg agattacaag gatgacgacg ataaggtttc aggaagcgga
     4021 gctactaact tcagcctgct gaagcaggct ggagacgtgg aggagaaccc tggacctgga
     4081 tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag
     4141 aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc
     4201 cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg
     4261 aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc
     4321 gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg
     4381 ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct
     4441 gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg
     4501 aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat
     4561 ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc
     4621 atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg
     4681 gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggccgaccgc
     4741 tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct
     4801 gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat
     4861 cgccttcttg acgagttctt ctgattcgaa catcacatta aatgagacct tttctgctat
     4921 tcaggaaata acccagttct gcaccactgg tttttgtagc tatctcgtaa ggctgctggc
     4981 tgaaaactgt gtctatgcaa ccttccaagt gcggagtgtc aaccaactgg acgggagaga
     5041 gtactgctcc tactccagga ctctcacaaa gctgatgagc tgtacttcag aaaaaaataa
     5101 taatttccat gttttgtata tatctgacaa aactggcaac atcttacaga ctactgactt
     5161 gaagacaacc tcttttatat ttctctattt ctgggctgat gaatttgttt tcatctgtct
     5221 tttccccctt cagaattttc cttggaaaaa aaatactagc ctagctggtc atttctttgt
     5281 aaggtagtta gcaattttaa gtctttcttt ggtcaacttt tttttaatgt gaaaagttag
     5341 gtaagacact tttttactgc ttttatgttt ttctgtcttg ttttgagacc atgatggtta
     5401 cacttttggt tcctaaataa aatttaaaaa attaacagcc aagtcacaaa ggtaatggat
     5461 tgcacataga ctaaggaata aacttcagat ttgtgatttt tgtttctaat cttgatgtaa
     5521 atttacacta tttataaata catatttatt gcttgaaaat atttgtgaat ggaatgctgt
     5581 tattttttcc agatttacct gccttggaaa gtcctctcca ctgactgtag cctccaattc
     5641 actggccgtc gttcactggc cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt
     5701 acccaactta atcgccttgc agcacatccc cctttcgcca gctggcgtaa tagcgaagag
     5761 gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg cgatttattc
     5821 aacaaagccg ccgtcccgtc aagtcagcgt aatgctctgc cagtgttaca accaattaac
     5881 caattctgat tagaaaaact catcgagcat caaatgaaac tgcaatatat tcatatcagg
     5941 attatcaata ccatattttt gaaaaagccg tttctgtaat gaaggagaaa actcaccgag
     6001 gcagttccat aggatggcaa gatcctggta tcggtctgcg attccgactc gtccaacatc
     6061 aatacaacct attaatttcc cctcgtcaaa aataaggtta tcaagtgaga aatcaccatg
     6121 agtgacgact gaatccggtg agaatggcaa aagcttatgc atttctttcc agacttgttc
     6181 aacaggccag ccattacgct cgtcatcaaa atcactcgca tcaaccaaac cgttattcat
     6241 tcgtgattgc gcctgagcga gacgaaatac gcgatcgctg ttaaaaggac aattacaaac
     6301 aggaatcgaa tgcaaccggc gcaggaacac tgccagcgca tcaacaatat tttcacctga
     6361 atcaggatat tcttctaata cctggaatgc tgttttcccg gggatcgcag tggtgagtaa
     6421 ccatgcatca tcaggagtac ggataaaatg cttgatggtc ggaagaggca taaattccgt
     6481 cagccagttt agtctgacca tctcatctgt aacatcattg gcaacgctac ctttgccatg
     6541 tttcagaaac aactctggcg catcgggctt cccatacaat cgatagattg tcgcacctga
     6601 ttgcccgaca ttatcgcgag cccatttata cccatataaa tcagcatcca tgttggaatt
     6661 taatcgcggc ttcgagcaag acgtttcccg ttgaatatgg ctcataacac cccttgtatt
     6721 actgtttatg taagcagaca gttttattgt tcatgatgat atatttttat cttgtgcaat
     6781 gtaacatcag agattttgag acacaacgtg gctttgttga ataaatcgaa cttttgctga
     6841 gttgaaggat cagatcacgc atcttcccga caacgcagac cgttccgtgg caaagcaaaa
     6901 gttcaaaatc accaactggt ccacctacaa caaagctctc atcaaccgtg gctccctcac
     6961 tttctggctg gatgatgggg cgattcaggc ctggtatgag tca
//
