LOCUS       Exported                7108 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Donor vector for 3' FLAG tag of human ZNF3.
ACCESSION   .
VERSION     .
KEYWORDS    pFETCh_ZNF3
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7108)
  TITLE     Mendenhall CRISPR tagging plasmids
REFERENCE   2  (bases 1 to 7108)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7108
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     CDS             791..814
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             818..841
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             845..868
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             884..940
                     /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             953..1747
                     /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(1007..1026)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1617..1636
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     CDS             2276..2287
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(2610..2627)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2610..2626)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2619..2641)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             complement(2845..3660)
                     /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     3568..3587
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3652..3671
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      4040..4628
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4529..4548
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4782..4799
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5099..5120
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5135..5165
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5173..5189
                     /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     5178..5200
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5197..5213
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5197..5213
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        5390..5592
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5543..5563
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     5589..5613
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             5689..6819
                     /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 gggagacaga ccccataagt gtgatgaatg tagcaagagc tttaatcgaa cttcagacct
       61 tattcaacat cagagaatcc acactgggga aaagccctat gaatgtaatg agtgtgggaa
      121 ggccttcagc cagagctcac accttattca gcatcagaga atccacactg gggaaaaacc
      181 ttatgaatgt agtgattgtg ggaaaacctt cagctgtagc tctgccctca ttctgcatcg
      241 gaggatccac acgggggaga aaccctatga atgtaatgag tgtgggaaga ccttcagctg
      301 gagctccacc ctcacccacc atcagagaat ccacactggt gagaaaccct acgcctgcaa
      361 tgaatgtggg aaggccttca gcaggagctc aacccttatt caccatcaga gaatccacac
      421 tggagaaaaa ccctatgaat gtaatgaatg tgggaaagcc ttcagccaga gctcacacct
      481 ctatcagcac cagagaatcc acactggaga gaagccctac gaatgtatgg aatgtggagg
      541 aaagtttacc tacagttcag gccttattca gcatcaaaga atccacaccg gggagaaccc
      601 ctatgaatgt agtgagtgtg ggaaagcctt caggtacagc tcggctcttg ttcgccatca
      661 gagaattcac actggagaga agcctttgaa tgggatcggc atgagcaaaa gctccctcag
      721 agttacgacc gagttaaata tcagagagtc cacggggagc ggaggaggtt ccggtggagg
      781 tggttctgga gattacaagg atgacgacga taagggcgat tacaaggatg acgacgataa
      841 gggagattac aaggatgacg acgataaggt ttcaggaagc ggagctacta acttcagcct
      901 gctgaagcag gctggagacg tggaggagaa ccctggacct ggatcgtttc gcatgattga
      961 acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga
     1021 ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg
     1081 gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga
     1141 ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt
     1201 tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct
     1261 gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct
     1321 gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg
     1381 agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca
     1441 ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga
     1501 tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt
     1561 ttctggattc atcgactgtg gccggctggg tgtggccgac cgctatcagg acatagcgtt
     1621 ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct
     1681 ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt
     1741 cttctgattc gaacatcacc cattttcctc actttccctg agtctcaaga gctcttgcct
     1801 taccctataa atctcaacag cttaggatgt gtccctttca actcagactt ttcattttag
     1861 agaatggggc agatggggca aatcgttgaa ttttcccaga aatcacacca gccttagaaa
     1921 gcgtcaaggc aagtggatgg cgtgctggga atagaaagca gctctgggac cagttacccc
     1981 atttaggaaa ggagtttgca ctaaactgtt tttctcacag cagaggaacc tttccaaggt
     2041 ggggatggaa gcacagtcgg gacagaattc gatggattcc tttagttgga gtccgcgtag
     2101 tcagcacaga cagcagtgga aaggacgctc tgggtcctgt tatttgctag ggagggtaaa
     2161 gggagactat ttcaaagcta ctgttcctag tccagcttta agtttcggta agaaacatgc
     2221 tgttttgttt catgatttcg ttaattatgg aaatttggca ttgagggatt attttattga
     2281 gggtagaaga gattccagaa tcatcatctg tgatgatggt gtcctttagg gctcttggag
     2341 cagccagacc atgtttccaa gagaaacctg gtgatattgc cagcagaccc cctgccatcc
     2401 cccccagttg tcctggggct gaatgggcaa atctgtccaa acagctagta accggctgtg
     2461 agggagaggg tcagaagcac ttagcgttgg cctctgattg ctgtcctctc ttgtcctctt
     2521 cccactccaa tgatgaaaat gattttctct aaatgccctt ggaaagtcct ctccactgac
     2581 tgtagcctcc aattcactgg ccgtcgttca ctggccgtcg ttttacaacg tcgtgactgg
     2641 gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt cgccagctgg
     2701 cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc
     2761 gaatgcgatt tattcaacaa agccgccgtc ccgtcaagtc agcgtaatgc tctgccagtg
     2821 ttacaaccaa ttaaccaatt ctgattagaa aaactcatcg agcatcaaat gaaactgcaa
     2881 tttattcata tcaggattat caataccata tttttgaaaa agccgtttct gtaatgaagg
     2941 agaaaactca ccgaggcagt tccataggat ggcaagatcc tggtatcggt ctgcgattcc
     3001 gactcgtcca acatcaatac aacctattaa tttcccctcg tcaaaaataa ggttatcaag
     3061 tgagaaatca ccatgagtga cgactgaatc cggtgagaat ggcaaaagct tatgcatttc
     3121 tttccagact tgttcaacag gccagccatt acgctcgtca tcaaaatcac tcgcatcaac
     3181 caaaccgtta ttcattcgtg attgcgcctg agcgagacga aatacgcgat cgctgttaaa
     3241 aggacaatta caaacaggaa tcgaatgcaa ccggcgcagg aacactgcca gcgcatcaac
     3301 aatattttca cctgaatcag gatattcttc taatacctgg aatgctgttt tcccggggat
     3361 cgcagtggtg agtaaccatg catcatcagg agtacggata aaatgcttga tggtcggaag
     3421 aggcataaat tccgtcagcc agtttagtct gaccatctca tctgtaacat cattggcaac
     3481 gctacctttg ccatgtttca gaaacaactc tggcgcatcg ggcttcccat acaatcgata
     3541 gattgtcgca cctgattgcc cgacattatc gcgagcccat ttatacccat ataaatcagc
     3601 atccatgttg gaatttaatc gcggcttcga gcaagacgtt tcccgttgaa tatggctcat
     3661 aacacccctt gtattactgt ttatgtaagc agacagtttt attgttcatg atgatatatt
     3721 tttatcttgt gcaatgtaac atcagagatt ttgagacaca acgtggcttt gttgaataaa
     3781 tcgaactttt gctgagttga aggatcagat cacgcatctt cccgacaacg cagaccgttc
     3841 cgtggcaaag caaaagttca aaatcaccaa ctggtccacc tacaacaaag ctctcatcaa
     3901 ccgtggctcc ctcactttct ggctggatga tggggcgatt caggcctggt atgagtcagc
     3961 aacaccttct tcacgaggca gacctctcga cggagttcca ctgagcgtca gaccccgtag
     4021 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     4081 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     4141 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
     4201 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     4261 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     4321 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     4381 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     4441 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     4501 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     4561 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     4621 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     4681 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg
     4741 agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg
     4801 aagcggaaga agctcgcaca ttcagcagcg tttttcagcg cgttttcgat caacgtttca
     4861 atgttggtat caacaccagg tttaactttg aacttatcgg cactgacggt tactgatttt
     4921 gaacttttgc tttgccacgg aacggtctgc gttgtcggga agatgcgtga tctgatcctt
     4981 caactcagca aaagttccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt
     5041 aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta
     5101 atgtgagtta gctcactcat taggcacccc aggctttaca ctttatgctt ccggctcgta
     5161 tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat gaccatgatt
     5221 acgccaagct tgcatgcctg caggtcgact ctagaggatc cccgggtacc gagctcgaat
     5281 tagcttgcat gcctgcaggt caattccctg gcattatgcc cagtacatga ccttatggga
     5341 ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg tgatgcggtt
     5401 ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc aagtctccac
     5461 cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt
     5521 cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat
     5581 ataagcagag ctcgtttagt gaaccgtcag atcgcctgga gacgccatcc acgctgtttt
     5641 gacctccata ggagacaccg ggaccgatcc agcctgggga tcccgaccat ggcttcgtac
     5701 ccctgccatc aacacgcgtc tgcgttcgac caggctgcgc gttctcgcgg ccatagcaac
     5761 cgacgtacgg cgttgcgccc tcgccggcag caagaagcca cggaagtccg cctggagcag
     5821 aaaatgccca cgctactgcg ggtttatata gacggtcctc acgggatggg gaaaaccacc
     5881 accacgcaac tgctggtggc cctgggttcg cgcgacgata tcgtctacgt acccgagccg
     5941 atgacttact ggcaggtgct gggggcttcc gagacaatcg cgaacatcta caccacacaa
     6001 caccgcctcg accagggtga gatatcggcc ggggacgcgg cggtggtaat gacaagcgcc
     6061 cagataacaa tgggcatgcc ttatgccgtg accgacgccg ttctggctcc tcatatcggg
     6121 ggggaggctg ggagctcaca tgccccgccc ccggccctca ccctcatctt cgaccgccat
     6181 cccatcgccg ccctcctgtg ctacccggcc gcgcgatacc ttatgggcag catgaccccc
     6241 caggccgtgc tggcgttcgt ggccctcatc ccgccgacct tgcccggcac aaacatcgtg
     6301 ttgggggccc ttccggagga cagacacatc gaccgcctgg ccaaacgcca gcgccccggc
     6361 gagcggcttg acctggctat gctggccgcg attcgccgcg tttacgggct gcttgccaat
     6421 acggtgcggt atctgcaggg cggcgggtcg tggcgggagg attggggaca gctttcgggg
     6481 acggccgtgc cgccccaggg tgccgagccc cagagcaacg cgggcccacg accccatatc
     6541 ggggacacgt tatttaccct gtttcgggcc cccgagttgc tggcccccaa cggcgacctg
     6601 tacaacgtgt ttgcctgggc cttggacgtc ttggccaaac gcctccgtcc catgcacgtc
     6661 tttatcctgg attacgacca atcgcccgcc ggctgccggg acgccctgct gcaacttacc
     6721 tccgggatgg tccagaccca cgtcaccacc cccggctcca taccgacgat ctgcgacctg
     6781 gcgcgcacgt ttgcccggga gatgggggag gctaactgag gatcccacgt cactattgta
     6841 tactctatat tatactctat gttatactct gtaatcctac tcaataaacg tgtcacgcct
     6901 gtgaaaccgt actaagtctc ccgtgtattc ttatcaccat caggtgacat cctcgcccag
     6961 gctgtcaatc atgccggtat cgattccagt agcaccggcc ccacgctgac aacccactct
     7021 tgcagcgtta gcagcgcccc tcttaacaag ccgaccccca ccagcgtcgc ggttactaac
     7081 actcctctcc ccgacctgca gcccagct
//
