LOCUS       sequence-199541-        6959 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor vector for 3' FLAG tag of human GABPB1_iso2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6959)
  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 6959)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Genome Res.
            2015 Sep 9."
COMMENT     Sequence Label: pFETCh_GABPB1_iso2
FEATURES             Location/Qualifiers
     source          1..6959
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(74..1204)
                     /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"
     primer_bind     complement(1280..1304)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        complement(1301..1503)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(1330..1350)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     complement(1680..1696)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1680..1696)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1693..1715)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1704..1720
                     /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)."
     promoter        complement(join(1728..1734,1735..1752,1753..1758))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1773..1794
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2094..2111)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2265..2853)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2345..2364)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(3222..3241)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3233..4048
                     /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     complement(3306..3325)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     4252..4274
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4267..4283
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(5043..5837)
                     /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(5154..5173)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5764..5783
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     CDS             complement(5850..5906)
                     /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             complement(5922..5945)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             complement(5949..5972)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             complement(5976..5999)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
ORIGIN
        1 cacgtttatt gagtaggatt acagagtata acatagagta taatatagag tatacaatag
       61 tgacgtggga tcctcagtta gcctccccca tctcccgggc aaacgtgcgc gccaggtcgc
      121 agatcgtcgg tatggagccg ggggtggtga cgtgggtctg gaccatcccg gaggtaagtt
      181 gcagcagggc gtcccggcag ccggcgggcg attggtcgta atccaggata aagacgtgca
      241 tgggacggag gcgtttggcc aagacgtcca aggcccaggc aaacacgttg tacaggtcgc
      301 cgttgggggc cagcaactcg ggggcccgaa acagggtaaa taacgtgtcc ccgatatggg
      361 gtcgtgggcc cgcgttgctc tggggctcgg caccctgggg cggcacggcc gtccccgaaa
      421 gctgtcccca atcctcccgc cacgacccgc cgccctgcag ataccgcacc gtattggcaa
      481 gcagcccgta aacgcggcga atcgcggcca gcatagccag gtcaagccgc tcgccggggc
      541 gctggcgttt ggccaggcgg tcgatgtgtc tgtcctccgg aagggccccc aacacgatgt
      601 ttgtgccggg caaggtcggc gggatgaggg ccacgaacgc cagcacggcc tggggggtca
      661 tgctgcccat aaggtatcgc gcggccgggt agcacaggag ggcggcgatg ggatggcggt
      721 cgaagatgag ggtgagggcc gggggcgggg catgtgagct cccagcctcc cccccgatat
      781 gaggagccag aacggcgtcg gtcacggcat aaggcatgcc cattgttatc tgggcgcttg
      841 tcattaccac cgccgcgtcc ccggccgata tctcaccctg gtcgaggcgg tgttgtgtgg
      901 tgtagatgtt cgcgattgtc tcggaagccc ccagcacctg ccagtaagtc atcggctcgg
      961 gtacgtagac gatatcgtcg cgcgaaccca gggccaccag cagttgcgtg gtggtggttt
     1021 tccccatccc gtgaggaccg tctatataaa cccgcagtag cgtgggcatt ttctgctcca
     1081 ggcggacttc cgtggcttct tgctgccggc gagggcgcaa cgccgtacgt cggttgctat
     1141 ggccgcgaga acgcgcagcc tggtcgaacg cagacgcgtg ttgatggcag gggtacgaag
     1201 ccatggtcgg gatccccagg ctggatcggt cccggtgtct cctatggagg tcaaaacagc
     1261 gtggatggcg tctccaggcg atctgacggt tcactaaacg agctctgctt atatagacct
     1321 cccaccgtac acgcctaccg cccatttgcg tcaatggggc ggagttgtta cgacattttg
     1381 gaaagtcccg ttgattttgg tgccaaaaca aactcccatt gacgtcaatg gggtggagac
     1441 ttgaaatccc cgtgagtcaa accgctatcc acgcccattg atgtactgcc aaaaccgcat
     1501 caccatggta atagcgatga ctaatacgta gatgtactgc caagtaggaa agtcccataa
     1561 ggtcatgtac tgggcataat gccagggaat tgacctgcag gcatgcaagc taattcgagc
     1621 tcggtacccg gggatcctct agagtcgacc tgcaggcatg caagcttggc gtaatcatgg
     1681 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
     1741 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
     1801 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     1861 ggccaacgcg cggggagagg cggtttgcgt attgggaact tttgctgagt tgaaggatca
     1921 gatcacgcat cttcccgaca acgcagaccg ttccgtggca aagcaaaagt tcaaaatcag
     1981 taaccgtcag tgccgataag ttcaaagtta aacctggtgt tgataccaac attgaaacgt
     2041 tgatcgaaaa cgcgctgaaa aacgctgctg aatgtgcgag cttcttccgc ttcctcgctc
     2101 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     2161 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     2221 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     2281 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     2341 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     2401 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     2461 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     2521 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     2581 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     2641 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     2701 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     2761 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     2821 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     2881 tctgacgctc agtggaactc cgtcgagagg tctgcctcgt gaagaaggtg ttgctgactc
     2941 ataccaggcc tgaatcgccc catcatccag ccagaaagtg agggagccac ggttgatgag
     3001 agctttgttg taggtggacc agttggtgat tttgaacttt tgctttgcca cggaacggtc
     3061 tgcgttgtcg ggaagatgcg tgatctgatc cttcaactca gcaaaagttc gatttattca
     3121 acaaagccac gttgtgtctc aaaatctctg atgttacatt gcacaagata aaaatatatc
     3181 atcatgaaca ataaaactgt ctgcttacat aaacagtaat acaaggggtg ttatgagcca
     3241 tattcaacgg gaaacgtctt gctcgaagcc gcgattaaat tccaacatgg atgctgattt
     3301 atatgggtat aaatgggctc gcgataatgt cgggcaatca ggtgcgacaa tctatcgatt
     3361 gtatgggaag cccgatgcgc cagagttgtt tctgaaacat ggcaaaggta gcgttgccaa
     3421 tgatgttaca gatgagatgg tcagactaaa ctggctgacg gaatttatgc ctcttccgac
     3481 catcaagcat tttatccgta ctcctgatga tgcatggtta ctcaccactg cgatccccgg
     3541 gaaaacagca ttccaggtat tagaagaata tcctgattca ggtgaaaata ttgttgatgc
     3601 gctggcagtg ttcctgcgcc ggttgcattc gattcctgtt tgtaattgtc cttttaacag
     3661 cgatcgcgta tttcgtctcg ctcaggcgca atcacgaatg aataacggtt tggttgatgc
     3721 gagtgatttt gatgacgagc gtaatggctg gcctgttgaa caagtctgga aagaaatgca
     3781 taagcttttg ccattctcac cggattcagt cgtcactcat ggtgatttct cacttgataa
     3841 ccttattttt gacgagggga aattaatagg ttgtattgat gttggacgag tcggaatcgc
     3901 agaccgatac caggatcttg ccatcctatg gaactgcctc ggtgagtttt ctccttcatt
     3961 acagaaacgg ctttttcaaa aatatggtat tgataatcct gatatgaata tattgcagtt
     4021 tcatttgatg ctcgatgagt ttttctaatc agaattggtt aattggttgt aacactggca
     4081 gagcattacg ctgacttgac gggacggcgg ctttgttgaa taaatcgcat tcgccattca
     4141 ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg
     4201 cgaaaggggg atgtgctgca aggcgattaa gttgggtaac gccagggttt tcccagtcac
     4261 gacgttgtaa aacgacggcc agtgaacgac ggccagtgaa ttggaggcta cagtcagtgg
     4321 agaggacttt ccaagctaac agagcaagag gtgggattcc aactcagttc atttgatttc
     4381 aagttcagtg gctgcagttg ttctttcaaa gttattcaag tataactttg ttttcatttg
     4441 atataactta ttatcaagtt catggaacaa aatttattta cagtcagtca agaaaaaaac
     4501 ctgtgaagca actatggttt tgttacacct acaaactggt ttacgcttac aaattacttt
     4561 gggagagggg aaaaaatgat gaatcctatg aacagctagt ctaaattcat ttacttgcct
     4621 tgaaataagc aattatctac tgcttttaaa aaataaccca tttgacaaac atcaacctcc
     4681 tttaagagga cccctagccc aattaggcat ttgagtttca gttttgaaca acaaaaaaaa
     4741 aattttagcc aatatatact ggtattttcc tttagtgacg tgtaagaatg gtgtgtgtaa
     4801 gtttaaaaca taaaggatga agtggaagaa ccaaagacat tccagtgtta tcaagatgac
     4861 agactgttat aaacaaattc ttcactcact cattcattca ttcaatattt atttattgag
     4921 ggcttgctat atgtataagg caaaaataaa caaaatgaaa atactacgca ctttcattct
     4981 gtatcaaaac aatactgtca gtaataaata taaaagacaa aaaaaattga atgatgttcg
     5041 aatcagaaga actcgtcaag aaggcgatag aaggcgatgc gctgcgaatc gggagcggcg
     5101 ataccgtaaa gcacgaggaa gcggtcagcc cattcgccgc caagctcttc agcaatatca
     5161 cgggtagcca acgctatgtc ctgatagcgg tcggccacac ccagccggcc acagtcgatg
     5221 aatccagaaa agcggccatt ttccaccatg atattcggca agcaggcatc gccatgggtc
     5281 acgacgagat cctcgccgtc gggcatgcgc gccttgagcc tggcgaacag ttcggctggc
     5341 gcgagcccct gatgctcttc gtccagatca tcctgatcga caagaccggc ttccatccga
     5401 gtacgtgctc gctcgatgcg atgtttcgct tggtggtcga atgggcaggt agccggatca
     5461 agcgtatgca gccgccgcat tgcatcagcc atgatggata ctttctcggc aggagcaagg
     5521 tgagatgaca ggagatcctg ccccggcact tcgcccaata gcagccagtc ccttcccgct
     5581 tcagtgacaa cgtcgagcac agctgcgcaa ggaacgcccg tcgtggccag ccacgatagc
     5641 cgcgctgcct cgtcctgcag ttcattcagg gcaccggaca ggtcggtctt gacaaaaaga
     5701 accgggcgcc cctgcgctga cagccggaac acggcggcat cagagcagcc gattgtctgt
     5761 tgtgcccagt catagccgaa tagcctctcc acccaagcgg ccggagaacc tgcgtgcaat
     5821 ccatcttgtt caatcatgcg aaacgatcca ggtccagggt tctcctccac gtctccagcc
     5881 tgcttcagca ggctgaagtt agtagctccg cttcctgaaa ccttatcgtc gtcatccttg
     5941 taatctccct tatcgtcgtc atccttgtaa tcgcccttat cgtcgtcatc cttgtaatct
     6001 ccagaaccac ctccaccgga acctcctccg ctccccttag ggtggctgcg gcaaagcaca
     6061 ccgggtaaaa gactccttac ttctatttct gcagattcca cccggttttc aattatttcg
     6121 atacattgtc tcttagctgg tggttcttca cttataacag tttcttcagc aatgtctgtt
     6181 gctggtactg ttaatactaa aatgaaaaaa aaattatgta ttacttcatt ttcagagaga
     6241 tttattagaa ataaaactag tcttgacatt gttgatgcta ctcatctaaa ataactacat
     6301 tccaaaacaa taattccaaa tcattgattt gctttttatg agactatcca tatttgtctt
     6361 ttttattacc ttcttgcttt tttactgaaa tctacttatt aaagtaccta caaatatttt
     6421 tttaattata tagataatat attcattcat gactcaaaac atcaaaatat ttaacgagca
     6481 tattttgaga ggcctcatgc ccacccctgc taacagatga tcattttatt aatttattat
     6541 gtattatcct tatattaatt tgtttaaaaa ttagtataca ttctcatttt cctttcttac
     6601 ataaaaggta gcatctatac aatagatata ttgttctgca tctgcttttt tcactccaaa
     6661 tacttataat atcttacatc actacataga gagcttctgt ttatttagct gcataatatc
     6721 cactaagtgc atattccatt atgagctggg ctgcaggtcg gggagaggag tgttagtaac
     6781 cgcgacgctg gtgggggtcg gcttgttaag aggggcgctg ctaacgctgc aagagtgggt
     6841 tgtcagcgtg gggccggtgc tactggaatc gataccggca tgattgacag cctgggcgag
     6901 gatgtcacct gatggtgata agaatacacg ggagacttag tacggtttca caggcgtga
//
