LOCUS       Exported                7154 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Donor vector for 3' FLAG tag of human HMG20A.
ACCESSION   .
VERSION     .
KEYWORDS    pFETCh_HMG20A
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7154)
  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 7154)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7154
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     rep_origin      57..645
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     546..565
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     799..816
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    1116..1137
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        1152..1182
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1190..1206
                     /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     1195..1217
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1214..1230
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1214..1230
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        1407..1609
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1560..1580
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     1606..1630
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             1706..2836
                     /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             3962..3985
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             3989..4012
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             4016..4039
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             4055..4111
                     /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             4124..4918
                     /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(4178..4197)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4788..4807
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5781..5798)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5781..5797)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5790..5812)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             complement(6016..6831)
                     /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     6739..6758
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     6823..6842
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
ORIGIN
        1 ctctcgacgg agttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
       61 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
      121 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
      181 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
      241 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
      301 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
      361 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
      421 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
      481 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
      541 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
      601 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
      661 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
      721 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
      781 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagaagc tcgcacattc
      841 agcagcgttt ttcagcgcgt tttcgatcaa cgtttcaatg ttggtatcaa caccaggttt
      901 aactttgaac ttatcggcac tgacggttac tgattttgaa cttttgcttt gccacggaac
      961 ggtctgcgtt gtcgggaaga tgcgtgatct gatccttcaa ctcagcaaaa gttcccaata
     1021 cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt
     1081 cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct cactcattag
     1141 gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga
     1201 taacaatttc acacaggaaa cagctatgac catgattacg ccaagcttgc atgcctgcag
     1261 gtcgactcta gaggatcccc gggtaccgag ctcgaattag cttgcatgcc tgcaggtcaa
     1321 ttccctggca ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct
     1381 acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg
     1441 gatagcggtt tgactcacgg ggatttcaag tctccacccc attgacgtca atgggagttt
     1501 gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac
     1561 gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctc gtttagtgaa
     1621 ccgtcagatc gcctggagac gccatccacg ctgttttgac ctccatagga gacaccggga
     1681 ccgatccagc ctggggatcc cgaccatggc ttcgtacccc tgccatcaac acgcgtctgc
     1741 gttcgaccag gctgcgcgtt ctcgcggcca tagcaaccga cgtacggcgt tgcgccctcg
     1801 ccggcagcaa gaagccacgg aagtccgcct ggagcagaaa atgcccacgc tactgcgggt
     1861 ttatatagac ggtcctcacg ggatggggaa aaccaccacc acgcaactgc tggtggccct
     1921 gggttcgcgc gacgatatcg tctacgtacc cgagccgatg acttactggc aggtgctggg
     1981 ggcttccgag acaatcgcga acatctacac cacacaacac cgcctcgacc agggtgagat
     2041 atcggccggg gacgcggcgg tggtaatgac aagcgcccag ataacaatgg gcatgcctta
     2101 tgccgtgacc gacgccgttc tggctcctca tatcgggggg gaggctggga gctcacatgc
     2161 cccgcccccg gccctcaccc tcatcttcga ccgccatccc atcgccgccc tcctgtgcta
     2221 cccggccgcg cgatacctta tgggcagcat gaccccccag gccgtgctgg cgttcgtggc
     2281 cctcatcccg ccgaccttgc ccggcacaaa catcgtgttg ggggcccttc cggaggacag
     2341 acacatcgac cgcctggcca aacgccagcg ccccggcgag cggcttgacc tggctatgct
     2401 ggccgcgatt cgccgcgttt acgggctgct tgccaatacg gtgcggtatc tgcagggcgg
     2461 cgggtcgtgg cgggaggatt ggggacagct ttcggggacg gccgtgccgc cccagggtgc
     2521 cgagccccag agcaacgcgg gcccacgacc ccatatcggg gacacgttat ttaccctgtt
     2581 tcgggccccc gagttgctgg cccccaacgg cgacctgtac aacgtgtttg cctgggcctt
     2641 ggacgtcttg gccaaacgcc tccgtcccat gcacgtcttt atcctggatt acgaccaatc
     2701 gcccgccggc tgccgggacg ccctgctgca acttacctcc gggatggtcc agacccacgt
     2761 caccaccccc ggctccatac cgacgatctg cgacctggcg cgcacgtttg cccgggagat
     2821 gggggaggct aactgaggat cccacgtcac tattgtatac tctatattat actctatgtt
     2881 atactctgta atcctactca ataaacgtgt cacgcctgtg aaaccgtact aagtctcccg
     2941 tgtattctta tcaccatcag gtgacatcct cgcccaggct gtcaatcatg ccggtatcga
     3001 ttccagtagc accggcccca cgctgacaac ccactcttgc agcgttagca gcgcccctct
     3061 taacaagccg acccccacca gcgtcgcggt tactaacact cctctccccg acctgcagcc
     3121 cagctaagtc ctcctgcctg agaactgtca gtgacagggg tgtgtgtgtt gtgtggagtg
     3181 gggtgctgtt tatgttagta ctggtgtgga gagattgaaa tctcctccag aggaaaatta
     3241 attaaactac attttttctt ctctggatat aatagaatgt cagtattggc taaaatcgtc
     3301 caatgtttca ggctctctgg tcattcaaat ttttgtccat ttatatttta atatctattt
     3361 tcccctcttt taagcttaat ttgctattca gaacactgga aactgagtag taaatcttga
     3421 gagtgtttct tcttatgtat gtttgtttgc tttttagttg tttttgtttt aacgggaagg
     3481 aagggagaat gaacactggt atttcttata tactctttca ttttgaagat agtcatagca
     3541 gaaagctgag cgagtctaac cttctgttac atgcagctgc tcatatatac tatacagcaa
     3601 ttatatagcc catctgttta tttagagtat gtggactaat ttcatttctt tgagatctaa
     3661 aagtctctcc tctaaaatta aaagcatgtc tgtggagcag accacattag aatcactgtt
     3721 tctttgaaat tgtgttttat ggtacaactg aatagggagg attttcttac tttcttctta
     3781 tctcacttca ggaagtggag agacacctac agtggacacc attgactcat atatgaacag
     3841 actgcacagt attattttag ctaatcccca agacaatgaa aacttcatag ctacagttcg
     3901 agaagttgtg aacagactcg atcgggggag cggaggaggt tccggtggag gtggttctgg
     3961 agattacaag gatgacgacg ataagggcga ttacaaggat gacgacgata agggagatta
     4021 caaggatgac gacgataagg tttcaggaag cggagctact aacttcagcc tgctgaagca
     4081 ggctggagac gtggaggaga accctggacc tggatcgttt cgcatgattg aacaagatgg
     4141 attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca
     4201 acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt
     4261 tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg
     4321 gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga
     4381 agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca
     4441 ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct
     4501 tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac
     4561 tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc
     4621 gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt
     4681 gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt
     4741 catcgactgt ggccggctgg gtgtggccga ccgctatcag gacatagcgt tggctacccg
     4801 tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat
     4861 cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgatt
     4921 cgaacatcga atggtgagtg ctcactgata aatatttata tgccagcaca tcatcaaaaa
     4981 taagatgtca tcagacttta tcaatactac taaaaccctg ggattacatt ggatgaacaa
     5041 gttggagact tggttaagat tcctgttgca tggttgttaa atgtagtaaa taatattaga
     5101 aaagagaatc actgtagtcc cagctacttg ggaggctgag gtgggaggat tgcttgagcc
     5161 cagcagttca agtccagaga gatcctgtct ctaaaaataa aagaaaacag actcaagaca
     5221 gttttttctt atgcttgctt cagaatggtt taatgggaaa agcatggttt ttgtggcttg
     5281 acctcatttc tagctttgtc acttattagc tgtattcttg ggcattagtt tcccaaaagg
     5341 ttttgaaaag tgagtgggaa acttagaaga aatcatcccc aaaacagcac tcagaacaaa
     5401 ataaatataa cttaattttt aatcttcctt ttctctggaa aatactaatg ataatttaga
     5461 agtagtcagt gagacattgt atgtatgtga gcctacataa gtaggaagct tttatattca
     5521 ccactcaata acacagtgac tgttgtctgt taaatgttga tgtggactca ccatacaaac
     5581 aggtaactcc ctggttttga tagattaaaa aataaactct tcagccaggt gcaatggctc
     5641 acacctgtaa gcctagcagt ttgggagttc aaggcaggca gatcgctgga gtccaggttc
     5701 aagaccagtc tgggcaacat gacaaaacct tggaaagtcc tctccactga ctgtagcctc
     5761 caattcactg gccgtcgttc actggccgtc gttttacaac gtcgtgactg ggaaaaccct
     5821 ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc
     5881 gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatgcgat
     5941 ttattcaaca aagccgccgt cccgtcaagt cagcgtaatg ctctgccagt gttacaacca
     6001 attaaccaat tctgattaga aaaactcatc gagcatcaaa tgaaactgca atatattcat
     6061 atcaggatta tcaataccat atttttgaaa aagccgtttc tgtaatgaag gagaaaactc
     6121 accgaggcag ttccatagga tggcaagatc ctggtatcgg tctgcgattc cgactcgtcc
     6181 aacatcaata caacctatta atttcccctc gtcaaaaata aggttatcaa gtgagaaatc
     6241 accatgagtg acgactgaat ccggtgagaa tggcaaaagc ttatgcattt ctttccagac
     6301 ttgttcaaca ggccagccat tacgctcgtc atcaaaatca ctcgcatcaa ccaaaccgtt
     6361 attcattcgt gattgcgcct gagcgagacg aaatacgcga tcgctgttaa aaggacaatt
     6421 acaaacagga atcgaatgca accggcgcag gaacactgcc agcgcatcaa caatattttc
     6481 acctgaatca ggatattctt ctaatacctg gaatgctgtt ttcccgggga tcgcagtggt
     6541 gagtaaccat gcatcatcag gagtacggat aaaatgcttg atggtcggaa gaggcataaa
     6601 ttccgtcagc cagtttagtc tgaccatctc atctgtaaca tcattggcaa cgctaccttt
     6661 gccatgtttc agaaacaact ctggcgcatc gggcttccca tacaatcgat agattgtcgc
     6721 acctgattgc ccgacattat cgcgagccca tttataccca tataaatcag catccatgtt
     6781 ggaatttaat cgcggcttcg agcaagacgt ttcccgttga atatggctca taacacccct
     6841 tgtattactg tttatgtaag cagacagttt tattgttcat gatgatatat ttttatcttg
     6901 tgcaatgtaa catcagagat tttgagacac aacgtggctt tgttgaataa atcgaacttt
     6961 tgctgagttg aaggatcaga tcacgcatct tcccgacaac gcagaccgtt ccgtggcaaa
     7021 gcaaaagttc aaaatcacca actggtccac ctacaacaaa gctctcatca accgtggctc
     7081 cctcactttc tggctggatg atggggcgat tcaggcctgg tatgagtcag caacaccttc
     7141 ttcacgaggc agac
//
