LOCUS       Exported                7193 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Donor vector for 3' FLAG tag of human SOX5.
ACCESSION   .
VERSION     .
KEYWORDS    pFETCh_SOX5
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7193)
  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 7193)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7193
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     CDS             837..860
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             864..887
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             891..914
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             930..986
                     /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             999..1793
                     /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(1053..1072)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1663..1682
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(2695..2712)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2695..2711)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2704..2726)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             complement(2930..3745)
                     /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     3653..3672
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3737..3756
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      4125..4713
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4614..4633
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4867..4884
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5184..5205
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5220..5250
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5258..5274
                     /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     5263..5285
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5282..5298
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5282..5298
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        5475..5677
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5628..5648
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     5674..5698
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             5774..6904
                     /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 acataaggta ttaggcctgg tgcggtggct cacgcctgta atcccaacac tttgggaggc
       61 cgaggcaggt ggatcacctg aggtcaggag ttcaagacca gcctggccaa catggtgaaa
      121 ccccgtctgt actaaaaata caaaagctag ccaggtgtgg tggcaggcgc ctgtattccc
      181 agctacttgg ggggctgagg caggagaatc gcttgaaccc tggaggggga ggttgcagtg
      241 aggcaagatc acgccattgc actgcagcct aggcaacagg agcgaaacta cgtcatctca
      301 aaaaaaaaaa aaaaaaagaa aagaaaagaa aaacaaggta ttaaagttca agtaggatgg
      361 agtttgaaaa atgttaggca attagataaa attagtacct tagaattgga cattgctgag
      421 caaattaatt atgtagtaaa gtccacatct aactattcac ttacccacga tgtctttttg
      481 gaaattcctt tcttgacagg caacaagcac agatccccat tgccactgct ggtgttgtgt
      541 accctggagc catcgccatg gctgggatgc cctcccctca cctgccctcg gagcactcaa
      601 gcgtgtctag cagcccagag cctgggatgc ctgttatcca gagcacttac ggtgtgaaag
      661 gagaggagcc acatatcaaa gaagagatac aggccgagga catcaatgga gaaatttatg
      721 atgagtacga cgaggaagag gatgatccag atgtagatta tgggagtgac agtgaaaacc
      781 atattgcagg acaagccaac gggagcggag gaggttccgg tggaggtggt tctggagatt
      841 acaaggatga cgacgataag ggcgattaca aggatgacga cgataaggga gattacaagg
      901 atgacgacga taaggtttca ggaagcggag ctactaactt cagcctgctg aagcaggctg
      961 gagacgtgga ggagaaccct ggacctggat cgtttcgcat gattgaacaa gatggattgc
     1021 acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga
     1081 caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt
     1141 ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat
     1201 cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg
     1261 gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg
     1321 ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc
     1381 cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga
     1441 tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag
     1501 ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc
     1561 atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg
     1621 actgtggccg gctgggtgtg gccgaccgct atcaggacat agcgttggct acccgtgata
     1681 ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg
     1741 ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc tgattcgaac
     1801 atcgggtcaa aagattgttg tgaccttagg acttaaagaa gccctaactg gttcatcctt
     1861 accagtggcc aagcacatta actttctcat acactgactg ttactttaac tgttagtctt
     1921 aaatagttgg gacatcagct gactaataga cctcagcctc aaaaggcttg gaaagaaaaa
     1981 acaaatacaa caagcaaaca acaatatcaa caacaagaga ttgaaataag ctatgggtaa
     2041 aataatgcca gtaattcagc tgctacatcc aagcactgaa gtcttacccg tcaacttttt
     2101 ttttttttta aataaacttt atggctgttt gttctacaat gttctagaaa ttctcactca
     2161 ggtacacagt gccaacaagt ggcttgtgaa tgtgttttgt tgttttgtgc tacaattttt
     2221 aaaaagaaaa aagttttgtt ttgttttttg gggtttctgg gttttttcct tttctttttc
     2281 tttcctttca ttttttttct ttgtaatgca cctgacagaa aaaaaagaaa aatgaatttc
     2341 tctttacttc tctccacctt ctccatctct ctactttaaa gatggaagtc tgtgcatgag
     2401 gggaaagagg gaaaaagagc ctgtttttaa cttccttgct atccaccaca aaataagcaa
     2461 ttattttctt tagaggactt tatctattgc acaccacact acatctttga gcaagtgcca
     2521 aatttgtact gaagtgttga ccaagttcat tttttctctt tactttttcc ttttccttct
     2581 taagttagga cagtgttaaa tcttagacaa tcccttgaaa aacctgaaat accagcagct
     2641 ggcttggaaa gtcctctcca ctgactgtag cctccaattc actggccgtc gttcactggc
     2701 cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt acccaactta atcgccttgc
     2761 agcacatccc cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc
     2821 ccaacagttg cgcagcctga atggcgaatg cgatttattc aacaaagccg ccgtcccgtc
     2881 aagtcagcgt aatgctctgc cagtgttaca accaattaac caattctgat tagaaaaact
     2941 catcgagcat caaatgaaac tgcaatttat tcatatcagg attatcaata ccatattttt
     3001 gaaaaagccg tttctgtaat gaaggagaaa actcaccgag gcagttccat aggatggcaa
     3061 gatcctggta tcggtctgcg attccgactc gtccaacatc aatacaacct attaatttcc
     3121 cctcgtcaaa aataaggtta tcaagtgaga aatcaccatg agtgacgact gaatccggtg
     3181 agaatggcaa aagcttatgc atttctttcc agacttgttc aacaggccag ccattacgct
     3241 cgtcatcaaa atcactcgca tcaaccaaac cgttattcat tcgtgattgc gcctgagcga
     3301 gacgaaatac gcgatcgctg ttaaaaggac aattacaaac aggaatcgaa tgcaaccggc
     3361 gcaggaacac tgccagcgca tcaacaatat tttcacctga atcaggatat tcttctaata
     3421 cctggaatgc tgttttcccg gggatcgcag tggtgagtaa ccatgcatca tcaggagtac
     3481 ggataaaatg cttgatggtc ggaagaggca taaattccgt cagccagttt agtctgacca
     3541 tctcatctgt aacatcattg gcaacgctac ctttgccatg tttcagaaac aactctggcg
     3601 catcgggctt cccatacaat cgatagattg tcgcacctga ttgcccgaca ttatcgcgag
     3661 cccatttata cccatataaa tcagcatcca tgttggaatt taatcgcggc ttcgagcaag
     3721 acgtttcccg ttgaatatgg ctcataacac cccttgtatt actgtttatg taagcagaca
     3781 gttttattgt tcatgatgat atatttttat cttgtgcaat gtaacatcag agattttgag
     3841 acacaacgtg gctttgttga ataaatcgaa cttttgctga gttgaaggat cagatcacgc
     3901 atcttcccga caacgcagac cgttccgtgg caaagcaaaa gttcaaaatc accaactggt
     3961 ccacctacaa caaagctctc atcaaccgtg gctccctcac tttctggctg gatgatgggg
     4021 cgattcaggc ctggtatgag tcagcaacac cttcttcacg aggcagacct ctcgacggag
     4081 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     4141 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     4201 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     4261 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     4321 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     4381 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     4441 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     4501 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     4561 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     4621 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     4681 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     4741 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     4801 gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc
     4861 gagcgcagcg agtcagtgag cgaggaagcg gaagaagctc gcacattcag cagcgttttt
     4921 cagcgcgttt tcgatcaacg tttcaatgtt ggtatcaaca ccaggtttaa ctttgaactt
     4981 atcggcactg acggttactg attttgaact tttgctttgc cacggaacgg tctgcgttgt
     5041 cgggaagatg cgtgatctga tccttcaact cagcaaaagt tcccaatacg caaaccgcct
     5101 ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa
     5161 gcgggcagtg agcgcaacgc aattaatgtg agttagctca ctcattaggc accccaggct
     5221 ttacacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata acaatttcac
     5281 acaggaaaca gctatgacca tgattacgcc aagcttgcat gcctgcaggt cgactctaga
     5341 ggatccccgg gtaccgagct cgaattagct tgcatgcctg caggtcaatt ccctggcatt
     5401 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
     5461 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
     5521 actcacgggg atttcaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca
     5581 aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg
     5641 taggcgtgta cggtgggagg tctatataag cagagctcgt ttagtgaacc gtcagatcgc
     5701 ctggagacgc catccacgct gttttgacct ccataggaga caccgggacc gatccagcct
     5761 ggggatcccg accatggctt cgtacccctg ccatcaacac gcgtctgcgt tcgaccaggc
     5821 tgcgcgttct cgcggccata gcaaccgacg tacggcgttg cgccctcgcc ggcagcaaga
     5881 agccacggaa gtccgcctgg agcagaaaat gcccacgcta ctgcgggttt atatagacgg
     5941 tcctcacggg atggggaaaa ccaccaccac gcaactgctg gtggccctgg gttcgcgcga
     6001 cgatatcgtc tacgtacccg agccgatgac ttactggcag gtgctggggg cttccgagac
     6061 aatcgcgaac atctacacca cacaacaccg cctcgaccag ggtgagatat cggccgggga
     6121 cgcggcggtg gtaatgacaa gcgcccagat aacaatgggc atgccttatg ccgtgaccga
     6181 cgccgttctg gctcctcata tcggggggga ggctgggagc tcacatgccc cgcccccggc
     6241 cctcaccctc atcttcgacc gccatcccat cgccgccctc ctgtgctacc cggccgcgcg
     6301 ataccttatg ggcagcatga ccccccaggc cgtgctggcg ttcgtggccc tcatcccgcc
     6361 gaccttgccc ggcacaaaca tcgtgttggg ggcccttccg gaggacagac acatcgaccg
     6421 cctggccaaa cgccagcgcc ccggcgagcg gcttgacctg gctatgctgg ccgcgattcg
     6481 ccgcgtttac gggctgcttg ccaatacggt gcggtatctg cagggcggcg ggtcgtggcg
     6541 ggaggattgg ggacagcttt cggggacggc cgtgccgccc cagggtgccg agccccagag
     6601 caacgcgggc ccacgacccc atatcgggga cacgttattt accctgtttc gggcccccga
     6661 gttgctggcc cccaacggcg acctgtacaa cgtgtttgcc tgggccttgg acgtcttggc
     6721 caaacgcctc cgtcccatgc acgtctttat cctggattac gaccaatcgc ccgccggctg
     6781 ccgggacgcc ctgctgcaac ttacctccgg gatggtccag acccacgtca ccacccccgg
     6841 ctccataccg acgatctgcg acctggcgcg cacgtttgcc cgggagatgg gggaggctaa
     6901 ctgaggatcc cacgtcacta ttgtatactc tatattatac tctatgttat actctgtaat
     6961 cctactcaat aaacgtgtca cgcctgtgaa accgtactaa gtctcccgtg tattcttatc
     7021 accatcaggt gacatcctcg cccaggctgt caatcatgcc ggtatcgatt ccagtagcac
     7081 cggccccacg ctgacaaccc actcttgcag cgttagcagc gcccctctta acaagccgac
     7141 ccccaccagc gtcgcggtta ctaacactcc tctccccgac ctgcagccca gct
//
