LOCUS       Exported                6892 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Donor vector for 3' FLAG tag of human ARID5B.
ACCESSION   .
VERSION     .
KEYWORDS    pFETCh_ARID5B
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6892)
  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 6892)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6892
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     rep_origin      40..628
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     529..548
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     782..799
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    1099..1120
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        1135..1165
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1173..1189
                     /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     1178..1200
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1197..1213
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1197..1213
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        1390..1592
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1543..1563
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     1589..1613
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             1689..2819
                     /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             3853..3876
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             3880..3903
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             3907..3930
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             3946..4002
                     /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             4015..4809
                     /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(4069..4088)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4679..4698
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5502..5519)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5502..5518)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5511..5533)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             complement(5737..6552)
                     /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     6460..6479
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     6544..6563
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
ORIGIN
        1 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
       61 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
      121 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
      181 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
      241 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
      301 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
      361 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
      421 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
      481 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
      541 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
      601 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
      661 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
      721 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
      781 cagcgagtca gtgagcgagg aagcggaaga agctcgcaca ttcagcagcg tttttcagcg
      841 cgttttcgat caacgtttca atgttggtat caacaccagg tttaactttg aacttatcgg
      901 cactgacggt tactgatttt gaacttttgc tttgccacgg aacggtctgc gttgtcggga
      961 agatgcgtga tctgatcctt caactcagca aaagttccca atacgcaaac cgcctctccc
     1021 cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg
     1081 cagtgagcgc aacgcaatta atgtgagtta gctcactcat taggcacccc aggctttaca
     1141 ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc ggataacaat ttcacacagg
     1201 aaacagctat gaccatgatt acgccaagct tgcatgcctg caggtcgact ctagaggatc
     1261 cccgggtacc gagctcgaat tagcttgcat gcctgcaggt caattccctg gcattatgcc
     1321 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
     1381 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
     1441 cggggatttc aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc
     1501 aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc
     1561 gtgtacggtg ggaggtctat ataagcagag ctcgtttagt gaaccgtcag atcgcctgga
     1621 gacgccatcc acgctgtttt gacctccata ggagacaccg ggaccgatcc agcctgggga
     1681 tcccgaccat ggcttcgtac ccctgccatc aacacgcgtc tgcgttcgac caggctgcgc
     1741 gttctcgcgg ccatagcaac cgacgtacgg cgttgcgccc tcgccggcag caagaagcca
     1801 cggaagtccg cctggagcag aaaatgccca cgctactgcg ggtttatata gacggtcctc
     1861 acgggatggg gaaaaccacc accacgcaac tgctggtggc cctgggttcg cgcgacgata
     1921 tcgtctacgt acccgagccg atgacttact ggcaggtgct gggggcttcc gagacaatcg
     1981 cgaacatcta caccacacaa caccgcctcg accagggtga gatatcggcc ggggacgcgg
     2041 cggtggtaat gacaagcgcc cagataacaa tgggcatgcc ttatgccgtg accgacgccg
     2101 ttctggctcc tcatatcggg ggggaggctg ggagctcaca tgccccgccc ccggccctca
     2161 ccctcatctt cgaccgccat cccatcgccg ccctcctgtg ctacccggcc gcgcgatacc
     2221 ttatgggcag catgaccccc caggccgtgc tggcgttcgt ggccctcatc ccgccgacct
     2281 tgcccggcac aaacatcgtg ttgggggccc ttccggagga cagacacatc gaccgcctgg
     2341 ccaaacgcca gcgccccggc gagcggcttg acctggctat gctggccgcg attcgccgcg
     2401 tttacgggct gcttgccaat acggtgcggt atctgcaggg cggcgggtcg tggcgggagg
     2461 attggggaca gctttcgggg acggccgtgc cgccccaggg tgccgagccc cagagcaacg
     2521 cgggcccacg accccatatc ggggacacgt tatttaccct gtttcgggcc cccgagttgc
     2581 tggcccccaa cggcgacctg tacaacgtgt ttgcctgggc cttggacgtc ttggccaaac
     2641 gcctccgtcc catgcacgtc tttatcctgg attacgacca atcgcccgcc ggctgccggg
     2701 acgccctgct gcaacttacc tccgggatgg tccagaccca cgtcaccacc cccggctcca
     2761 taccgacgat ctgcgacctg gcgcgcacgt ttgcccggga gatgggggag gctaactgag
     2821 gatcccacgt cactattgta tactctatat tatactctat gttatactct gtaatcctac
     2881 tcaataaacg tgtcacgcct gtgaaaccgt actaagtctc ccgtgtattc ttatcaccat
     2941 caggtgacat cctcgcccag gctgtcaatc atgccggtat cgattccagt agcaccggcc
     3001 ccacgctgac aacccactct tgcagcgtta gcagcgcccc tcttaacaag ccgaccccca
     3061 ccagcgtcgc ggttactaac actcctctcc ccgacctgca gcccagctaa agcctgtcgg
     3121 gtatcaccca tgaccatgtc aggccctaaa aaataccctg aatcgctttc aagatcagga
     3181 aaacctcacc atgtgagact ggagaatttc aggaagatgg aaggcatggt ccacccaatc
     3241 ctgcaccgga aaatgagccc gcagaacatt ggggcggcgc ggccgatcaa gcgcagcctg
     3301 gaggatttgg accttgtgat tgcagggaaa aaggcccggg cagtgtctcc cttagaccca
     3361 tccaaggagg tctctgggaa ggagaaggcc tctgagcagg agagtgaagg cagcaaagca
     3421 gcgcacggtg ggcattccgg gggcggatca gaaggccaca agcttcccct ctcctcccct
     3481 atcttcccag gtctgtattc cgggagcctg tgtaactcgg gcctcaactc caggctcccg
     3541 gctgggtatt ctcattctct gcagtacttg aaaaaccaga ctgtgctttc tccactcatg
     3601 cagcccctgg ctttccactc gcttgtgatg caaagaggaa tttttacatc accgacaaat
     3661 tctcagcagc tgtacagaca cttggctgcg gctacacctg taggaagttc atatggggac
     3721 cttttgcata acagcattta ccctttagct gctataaatc ctcaagctgc ctttccatct
     3781 tcccagctgt catccgtgca ccccagtact aagctcggga gcggaggagg ttccggtgga
     3841 ggtggttctg gagattacaa ggatgacgac gataagggcg attacaagga tgacgacgat
     3901 aagggagatt acaaggatga cgacgataag gtttcaggaa gcggagctac taacttcagc
     3961 ctgctgaagc aggctggaga cgtggaggag aaccctggac ctggatcgtt tcgcatgatt
     4021 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat
     4081 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag
     4141 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac
     4201 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac
     4261 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc
     4321 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg
     4381 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag
     4441 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat
     4501 caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag
     4561 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc
     4621 ttttctggat tcatcgactg tggccggctg ggtgtggccg accgctatca ggacatagcg
     4681 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg
     4741 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag
     4801 ttcttctgat tcgaacatcg ctcagctctg cccagcagtc caaagcggca tggccaacag
     4861 agcttcactc cttacccagg agtgctggct tatagagtta gaagtcagta tttcttctaa
     4921 tctgaggcta tgatcagtcc cagctgtagg ggcccagagg ggaggtgaac atgcctgatt
     4981 tttgtgggac aactctagcc cacaaactga ctggctggtg agtcttgact cccttccaac
     5041 acagatgccc aggcacctcc agatcattca cttcgcacgt gggccttgtg aagggatttg
     5101 tgaatatcca ggaagaactt agaggacccc atctgagttc ggatggtcag gaaacaatct
     5161 gggcaaaaaa gaggcaggca tttcaaagga aggggcaagg aagactggca aacagatggc
     5221 aagggatgcc cctctttttc ataaaactct ccaaggttca atcaatgcaa tgtatagtga
     5281 aacttcaata gatctttcat tttgacacta ttaaacaatc cagagaagta aacactgtta
     5341 aattgactgt atatatttgc ttcttaaaac tacctgtatc actgtttgct cacctaattt
     5401 atatacaggt agttccattt tctcccagtt ccttctcgtc tttttttttc ttggaaagtc
     5461 ctctccactg actgtagcct ccaattcact ggccgtcgtt cactggccgt cgttttacaa
     5521 cgtcgtgact gggaaaaccc tggcgttacc caacttaatc gccttgcagc acatccccct
     5581 ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc
     5641 agcctgaatg gcgaatgcga tttattcaac aaagccgccg tcccgtcaag tcagcgtaat
     5701 gctctgccag tgttacaacc aattaaccaa ttctgattag aaaaactcat cgagcatcaa
     5761 atgaaactgc aatatattca tatcaggatt atcaatacca tatttttgaa aaagccgttt
     5821 ctgtaatgaa ggagaaaact caccgaggca gttccatagg atggcaagat cctggtatcg
     5881 gtctgcgatt ccgactcgtc caacatcaat acaacctatt aatttcccct cgtcaaaaat
     5941 aaggttatca agtgagaaat caccatgagt gacgactgaa tccggtgaga atggcaaaag
     6001 cttatgcatt tctttccaga cttgttcaac aggccagcca ttacgctcgt catcaaaatc
     6061 actcgcatca accaaaccgt tattcattcg tgattgcgcc tgagcgagac gaaatacgcg
     6121 atcgctgtta aaaggacaat tacaaacagg aatcgaatgc aaccggcgca ggaacactgc
     6181 cagcgcatca acaatatttt cacctgaatc aggatattct tctaatacct ggaatgctgt
     6241 tttcccgggg atcgcagtgg tgagtaacca tgcatcatca ggagtacgga taaaatgctt
     6301 gatggtcgga agaggcataa attccgtcag ccagtttagt ctgaccatct catctgtaac
     6361 atcattggca acgctacctt tgccatgttt cagaaacaac tctggcgcat cgggcttccc
     6421 atacaatcga tagattgtcg cacctgattg cccgacatta tcgcgagccc atttataccc
     6481 atataaatca gcatccatgt tggaatttaa tcgcggcttc gagcaagacg tttcccgttg
     6541 aatatggctc ataacacccc ttgtattact gtttatgtaa gcagacagtt ttattgttca
     6601 tgatgatata tttttatctt gtgcaatgta acatcagaga ttttgagaca caacgtggct
     6661 ttgttgaata aatcgaactt ttgctgagtt gaaggatcag atcacgcatc ttcccgacaa
     6721 cgcagaccgt tccgtggcaa agcaaaagtt caaaatcacc aactggtcca cctacaacaa
     6781 agctctcatc aaccgtggct ccctcacttt ctggctggat gatggggcga ttcaggcctg
     6841 gtatgagtca gcaacacctt cttcacgagg cagacctctc gacggagttc ca
//
