LOCUS       Exported                6751 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Donor vector for 3' FLAG tag of human GATAD2A.
ACCESSION   .
VERSION     .
KEYWORDS    pFETCh_GATAD2A
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6751)
  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 6751)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6751
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     CDS             729..752
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             756..779
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             783..806
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             822..878
                     /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             891..1685
                     /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(945..964)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1555..1574
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(2240..2257)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2240..2256)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2249..2271)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             complement(2475..3290)
                     /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     3198..3217
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3282..3301
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      3670..4258
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4159..4178
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4412..4429
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4729..4750
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4765..4795
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4803..4819
                     /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     4808..4830
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4827..4843
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4827..4843
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        5020..5222
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5173..5193
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     5219..5243
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             5319..6449
                     /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 taactgggac tacagggatg tgccaccatg tccagctgat atttttttag agatggggcc
       61 tcactatgtt gcccaggctg gtctctatct cctggcctca agcagtcttc ccacccccca
      121 gcctctcaaa gtgctgggat tcaggctgag cctccgtgcc cagctaagat agtttttcat
      181 tgctgcatag tattccatcc tgcattattg tcctgttgca cacatttgtt tatggttttc
      241 cactgttcta agcagcaccg tggtcagcac ttcatttctg tagaacgtag gtagctgctc
      301 accctctgat gattgcccca cacttgggat taccttggca gagcctggca atgtcaaggc
      361 cattgagttg ctgcctaaat actcttcagc tctggggggg cctgggattt gaaacgttga
      421 ttgaggagta gtgtgggtgg gtttgcaagg gctcctgggt ccttagtctg tattggggct
      481 ggggtctagg cagctatggc tgggctcctc ccaggagccc tcctgacgag ggccttctca
      541 gctggtcgct ctgttctgtt gcaggcggga cccttgcgtt tgtcagccca agcctggcgg
      601 tgcacaagag ctcctcggcc gtggaccgcc agcgagagta cctcctggac atgatcccac
      661 cccgctccat cccccagtca gccacgtgga aagggagcgg aggaggttcc ggtggaggtg
      721 gttctggaga ttacaaggat gacgacgata agggcgatta caaggatgac gacgataagg
      781 gagattacaa ggatgacgac gataaggttt caggaagcgg agctactaac ttcagcctgc
      841 tgaagcaggc tggagacgtg gaggagaacc ctggacctgg atcgtttcgc atgattgaac
      901 aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact
      961 gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc
     1021 gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg
     1081 cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg
     1141 tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt
     1201 catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc
     1261 atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag
     1321 cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg
     1381 ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc
     1441 tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt
     1501 ctggattcat cgactgtggc cggctgggtg tggccgaccg ctatcaggac atagcgttgg
     1561 ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt
     1621 acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct
     1681 tctgattcga acatcggaag acgggctccc tcctccccca cctggcccct ggtctagaag
     1741 gacccactgc accaccctcc gctggctcgg gaagacaccg tgcccgcccc aagagcaagc
     1801 accggccatg ctgcagaggc aagacctcaa ttcttggctg caaagtttca tcagggctag
     1861 ggggctggtg ccgcctcata ggcagacgag gatcatcgct gggggacctt tcccgtgggc
     1921 tttcttcctt tctctctttg cctttagttt gcccgacacc agcagaaaag tggaccttgg
     1981 gggctggttc tgctcctggc ccccttgttc agcccctgcc ggcacacggg cggctcaccc
     2041 tggacactgt gatgcgcatg ggcaaggcca gcgcccgggg cttctgaacc gagcggggtg
     2101 tttcattttt ttgcttttcc ctgtcttagg ctcccagtct ttgactgcct tcccatggcg
     2161 atctataagt tgaaagattt tttttttctt ggaaagtcct ctccactgac tgtagcctcc
     2221 aattcactgg ccgtcgttca ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg
     2281 gcgttaccca acttaatcgc cttgcagcac atcccccttt cgccagctgg cgtaatagcg
     2341 aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc gaatgcgatt
     2401 tattcaacaa agccgccgtc ccgtcaagtc agcgtaatgc tctgccagtg ttacaaccaa
     2461 ttaaccaatt ctgattagaa aaactcatcg agcatcaaat gaaactgcaa tatattcata
     2521 tcaggattat caataccata tttttgaaaa agccgtttct gtaatgaagg agaaaactca
     2581 ccgaggcagt tccataggat ggcaagatcc tggtatcggt ctgcgattcc gactcgtcca
     2641 acatcaatac aacctattaa tttcccctcg tcaaaaataa ggttatcaag tgagaaatca
     2701 ccatgagtga cgactgaatc cggtgagaat ggcaaaagct tatgcatttc tttccagact
     2761 tgttcaacag gccagccatt acgctcgtca tcaaaatcac tcgcatcaac caaaccgtta
     2821 ttcattcgtg attgcgcctg agcgagacga aatacgcgat cgctgttaaa aggacaatta
     2881 caaacaggaa tcgaatgcaa ccggcgcagg aacactgcca gcgcatcaac aatattttca
     2941 cctgaatcag gatattcttc taatacctgg aatgctgttt tcccggggat cgcagtggtg
     3001 agtaaccatg catcatcagg agtacggata aaatgcttga tggtcggaag aggcataaat
     3061 tccgtcagcc agtttagtct gaccatctca tctgtaacat cattggcaac gctacctttg
     3121 ccatgtttca gaaacaactc tggcgcatcg ggcttcccat acaatcgata gattgtcgca
     3181 cctgattgcc cgacattatc gcgagcccat ttatacccat ataaatcagc atccatgttg
     3241 gaatttaatc gcggcttcga gcaagacgtt tcccgttgaa tatggctcat aacacccctt
     3301 gtattactgt ttatgtaagc agacagtttt attgttcatg atgatatatt tttatcttgt
     3361 gcaatgtaac atcagagatt ttgagacaca acgtggcttt gttgaataaa tcgaactttt
     3421 gctgagttga aggatcagat cacgcatctt cccgacaacg cagaccgttc cgtggcaaag
     3481 caaaagttca aaatcaccaa ctggtccacc tacaacaaag ctctcatcaa ccgtggctcc
     3541 ctcactttct ggctggatga tggggcgatt caggcctggt atgagtcagc aacaccttct
     3601 tcacgaggca gacctctcga cggagttcca ctgagcgtca gaccccgtag aaaagatcaa
     3661 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     3721 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     3781 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     3841 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     3901 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     3961 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     4021 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     4081 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     4141 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     4201 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     4261 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
     4321 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
     4381 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
     4441 agctcgcaca ttcagcagcg tttttcagcg cgttttcgat caacgtttca atgttggtat
     4501 caacaccagg tttaactttg aacttatcgg cactgacggt tactgatttt gaacttttgc
     4561 tttgccacgg aacggtctgc gttgtcggga agatgcgtga tctgatcctt caactcagca
     4621 aaagttccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg
     4681 gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta
     4741 gctcactcat taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg
     4801 aattgtgagc ggataacaat ttcacacagg aaacagctat gaccatgatt acgccaagct
     4861 tgcatgcctg caggtcgact ctagaggatc cccgggtacc gagctcgaat tagcttgcat
     4921 gcctgcaggt caattccctg gcattatgcc cagtacatga ccttatggga ctttcctact
     4981 tggcagtaca tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac
     5041 atcaatgggc gtggatagcg gtttgactca cggggatttc aagtctccac cccattgacg
     5101 tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact
     5161 ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag
     5221 ctcgtttagt gaaccgtcag atcgcctgga gacgccatcc acgctgtttt gacctccata
     5281 ggagacaccg ggaccgatcc agcctgggga tcccgaccat ggcttcgtac ccctgccatc
     5341 aacacgcgtc tgcgttcgac caggctgcgc gttctcgcgg ccatagcaac cgacgtacgg
     5401 cgttgcgccc tcgccggcag caagaagcca cggaagtccg cctggagcag aaaatgccca
     5461 cgctactgcg ggtttatata gacggtcctc acgggatggg gaaaaccacc accacgcaac
     5521 tgctggtggc cctgggttcg cgcgacgata tcgtctacgt acccgagccg atgacttact
     5581 ggcaggtgct gggggcttcc gagacaatcg cgaacatcta caccacacaa caccgcctcg
     5641 accagggtga gatatcggcc ggggacgcgg cggtggtaat gacaagcgcc cagataacaa
     5701 tgggcatgcc ttatgccgtg accgacgccg ttctggctcc tcatatcggg ggggaggctg
     5761 ggagctcaca tgccccgccc ccggccctca ccctcatctt cgaccgccat cccatcgccg
     5821 ccctcctgtg ctacccggcc gcgcgatacc ttatgggcag catgaccccc caggccgtgc
     5881 tggcgttcgt ggccctcatc ccgccgacct tgcccggcac aaacatcgtg ttgggggccc
     5941 ttccggagga cagacacatc gaccgcctgg ccaaacgcca gcgccccggc gagcggcttg
     6001 acctggctat gctggccgcg attcgccgcg tttacgggct gcttgccaat acggtgcggt
     6061 atctgcaggg cggcgggtcg tggcgggagg attggggaca gctttcgggg acggccgtgc
     6121 cgccccaggg tgccgagccc cagagcaacg cgggcccacg accccatatc ggggacacgt
     6181 tatttaccct gtttcgggcc cccgagttgc tggcccccaa cggcgacctg tacaacgtgt
     6241 ttgcctgggc cttggacgtc ttggccaaac gcctccgtcc catgcacgtc tttatcctgg
     6301 attacgacca atcgcccgcc ggctgccggg acgccctgct gcaacttacc tccgggatgg
     6361 tccagaccca cgtcaccacc cccggctcca taccgacgat ctgcgacctg gcgcgcacgt
     6421 ttgcccggga gatgggggag gctaactgag gatcccacgt cactattgta tactctatat
     6481 tatactctat gttatactct gtaatcctac tcaataaacg tgtcacgcct gtgaaaccgt
     6541 actaagtctc ccgtgtattc ttatcaccat caggtgacat cctcgcccag gctgtcaatc
     6601 atgccggtat cgattccagt agcaccggcc ccacgctgac aacccactct tgcagcgtta
     6661 gcagcgcccc tcttaacaag ccgaccccca ccagcgtcgc ggttactaac actcctctcc
     6721 ccgacctgca gcccagcttc agcctctgga g
//
