LOCUS       sequence-271142-        6970 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Donor vector for 3' FLAG tag of human MAF1_iso1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6970)
  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 6970)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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_MAF1_iso1
FEATURES             Location/Qualifiers
     source          1..6970
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..203
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     154..174
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     200..224
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             300..1430
                     /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             2464..2487
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2491..2514
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2518..2541
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             2557..2613
                     /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             2626..3420
                     /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(2680..2699)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3290..3309
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4191..4207)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4200..4222)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             complement(4426..5241)
                     /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     5149..5168
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     5233..5252
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      5621..6209
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6110..6129
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6363..6380
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6680..6701
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6716..6721,6722..6739,6740..6746)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6754..6770
                     /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     6759..6781
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6778..6794
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6778..6794
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
       61 caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact
      121 ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt
      181 gggaggtcta tataagcaga gctcgtttag tgaaccgtca gatcgcctgg agacgccatc
      241 cacgctgttt tgacctccat aggagacacc gggaccgatc cagcctgggg atcccgacca
      301 tggcttcgta cccctgccat caacacgcgt ctgcgttcga ccaggctgcg cgttctcgcg
      361 gccatagcaa ccgacgtacg gcgttgcgcc ctcgccggca gcaagaagcc acggaagtcc
      421 gcctggagca gaaaatgccc acgctactgc gggtttatat agacggtcct cacgggatgg
      481 ggaaaaccac caccacgcaa ctgctggtgg ccctgggttc gcgcgacgat atcgtctacg
      541 tacccgagcc gatgacttac tggcaggtgc tgggggcttc cgagacaatc gcgaacatct
      601 acaccacaca acaccgcctc gaccagggtg agatatcggc cggggacgcg gcggtggtaa
      661 tgacaagcgc ccagataaca atgggcatgc cttatgccgt gaccgacgcc gttctggctc
      721 ctcatatcgg gggggaggct gggagctcac atgccccgcc cccggccctc accctcatct
      781 tcgaccgcca tcccatcgcc gccctcctgt gctacccggc cgcgcgatac cttatgggca
      841 gcatgacccc ccaggccgtg ctggcgttcg tggccctcat cccgccgacc ttgcccggca
      901 caaacatcgt gttgggggcc cttccggagg acagacacat cgaccgcctg gccaaacgcc
      961 agcgccccgg cgagcggctt gacctggcta tgctggccgc gattcgccgc gtttacgggc
     1021 tgcttgccaa tacggtgcgg tatctgcagg gcggcgggtc gtggcgggag gattggggac
     1081 agctttcggg gacggccgtg ccgccccagg gtgccgagcc ccagagcaac gcgggcccac
     1141 gaccccatat cggggacacg ttatttaccc tgtttcgggc ccccgagttg ctggccccca
     1201 acggcgacct gtacaacgtg tttgcctggg ccttggacgt cttggccaaa cgcctccgtc
     1261 ccatgcacgt ctttatcctg gattacgacc aatcgcccgc cggctgccgg gacgccctgc
     1321 tgcaacttac ctccgggatg gtccagaccc acgtcaccac ccccggctcc ataccgacga
     1381 tctgcgacct ggcgcgcacg tttgcccggg agatggggga ggctaactga ggatcccacg
     1441 tcactattgt atactctata ttatactcta tgttatactc tgtaatccta ctcaataaac
     1501 gtgtcacgcc tgtgaaaccg tactaagtct cccgtgtatt cttatcacca tcaggtgaca
     1561 tcctcgccca ggctgtcaat catgccggta tcgattccag tagcaccggc cccacgctga
     1621 caacccactc ttgcagcgtt agcagcgccc ctcttaacaa gccgaccccc accagcgtcg
     1681 cggttactaa cactcctctc cccgacctgc agcccagctt ctgaaaccac agctgtggaa
     1741 cgcggtggac gaggagatct gcctggctga atgtgacatc tacaggtggg ccggcatccc
     1801 tgcagatggc ctccaggcct ccaggtttgt ggggcacttg gcgcctcaca ccacctgtca
     1861 ccctatactc cccagctata acccagactt ggactcagat cccttcgggg aggatggtag
     1921 cctctggtcc ttcaactact tcttctacaa caagcggctc aagcgaatcg tcttctttag
     1981 ctgccgttcc atcaggtggg caccccccgc ctcctccccc acctccctgt tgggccgata
     2041 caacttcccc agctgtgcca tctccaccca gctttgccct ccctgaacat cctccctggg
     2101 gccagcagca ggcctgggta ggggtcctga aactggtttc cctgcctccc ctcagtggct
     2161 ccacctacac accctcagag gcaggcaacg agctggacat ggagctgggg gaggaggagg
     2221 tggaggaaga aagcagaagc gggggcagtg gggccgagga gaccagcacc atggaggagg
     2281 acaggtgtgt gatgggggcc atgacccggg tccttgaagc ctggagtggg tacaacaggg
     2341 tgggggcctc ctctacttgg tctaagtggt ggctcctgaa ggcccactgt gtgccaacct
     2401 ccgcccaggg tcccagtgat ctgtattggg agcggaggag gttccggtgg aggtggttct
     2461 ggagattaca aggatgacga cgataagggc gattacaagg atgacgacga taagggagat
     2521 tacaaggatg acgacgataa ggtttcagga agcggagcta ctaacttcag cctgctgaag
     2581 caggctggag acgtggagga gaaccctgga cctggatcgt ttcgcatgat tgaacaagat
     2641 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca
     2701 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg
     2761 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg
     2821 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact
     2881 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct
     2941 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg
     3001 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt
     3061 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc
     3121 gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc
     3181 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga
     3241 ttcatcgact gtggccggct gggtgtggcc gaccgctatc aggacatagc gttggctacc
     3301 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt
     3361 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga
     3421 ttcgaacatc ccgaggcccc agcttcatcc agcttcaacc aatgcctgga cctgtccacc
     3481 tgagaggccc ctggggcctc cccagctgct ggccagaccc tggcgctgcc acagtcctgg
     3541 cactgcccaa ggccatacct gcctagccct ttggctccat cctgtggatg cccactcacc
     3601 cctcagactc ctgctgccca tgctgtggcc ggacttgtca gcagggggcc tggtgggagg
     3661 agcgactgcc ctgcccaaat gaactgccac agcagggaca gctggaccgc agagtttatt
     3721 tttgtatttc tactgggcct gcacactcca gcccaaaggg tctgtggccg gaggccccac
     3781 gagcaggccc cagcagtcac cggctctggt cttgggccgg ccccggtgcc cacctgtacc
     3841 cccacctcgc ccatttggcc gcgtgcactg agtgtcactt tgctgcagct cgtttctttc
     3901 caataaaagt ttctgtgact tagtgtggac ctgatgcggt gtgggggtgg ggcctgcggt
     3961 gtgggagtgg ccggcggcat ctggtatcct agcaactgcc gctgcgggcc gtgaccgcta
     4021 agtcctggga gctgttgggc tctgggaccg cagttgctga aatggaggca gaggtgtggg
     4081 aggcagaagg ctacaaccta gttctggact cggacctgta tgatgcggat ggctatgact
     4141 tggaaagtcc tctccactga ctgtagcctc caattcactg gccgtcgttc actggccgtc
     4201 gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca
     4261 catccccctt tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa
     4321 cagttgcgca gcctgaatgg cgaatgcgat ttattcaaca aagccgccgt cccgtcaagt
     4381 cagcgtaatg ctctgccagt gttacaacca attaaccaat tctgattaga aaaactcatc
     4441 gagcatcaaa tgaaactgca atatattcat atcaggatta tcaataccat atttttgaaa
     4501 aagccgtttc tgtaatgaag gagaaaactc accgaggcag ttccatagga tggcaagatc
     4561 ctggtatcgg tctgcgattc cgactcgtcc aacatcaata caacctatta atttcccctc
     4621 gtcaaaaata aggttatcaa gtgagaaatc accatgagtg acgactgaat ccggtgagaa
     4681 tggcaaaagc ttatgcattt ctttccagac ttgttcaaca ggccagccat tacgctcgtc
     4741 atcaaaatca ctcgcatcaa ccaaaccgtt attcattcgt gattgcgcct gagcgagacg
     4801 aaatacgcga tcgctgttaa aaggacaatt acaaacagga atcgaatgca accggcgcag
     4861 gaacactgcc agcgcatcaa caatattttc acctgaatca ggatattctt ctaatacctg
     4921 gaatgctgtt ttcccgggga tcgcagtggt gagtaaccat gcatcatcag gagtacggat
     4981 aaaatgcttg atggtcggaa gaggcataaa ttccgtcagc cagtttagtc tgaccatctc
     5041 atctgtaaca tcattggcaa cgctaccttt gccatgtttc agaaacaact ctggcgcatc
     5101 gggcttccca tacaatcgat agattgtcgc acctgattgc ccgacattat cgcgagccca
     5161 tttataccca tataaatcag catccatgtt ggaatttaat cgcggcttcg agcaagacgt
     5221 ttcccgttga atatggctca taacacccct tgtattactg tttatgtaag cagacagttt
     5281 tattgttcat gatgatatat ttttatcttg tgcaatgtaa catcagagat tttgagacac
     5341 aacgtggctt tgttgaataa atcgaacttt tgctgagttg aaggatcaga tcacgcatct
     5401 tcccgacaac gcagaccgtt ccgtggcaaa gcaaaagttc aaaatcacca actggtccac
     5461 ctacaacaaa gctctcatca accgtggctc cctcactttc tggctggatg atggggcgat
     5521 tcaggcctgg tatgagtcag caacaccttc ttcacgaggc agacctctcg acggagttcc
     5581 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     5641 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     5701 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     5761 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     5821 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     5881 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     5941 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     6001 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     6061 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     6121 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     6181 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     6241 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
     6301 ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc
     6361 gcagcgagtc agtgagcgag gaagcggaag aagctcgcac attcagcagc gtttttcagc
     6421 gcgttttcga tcaacgtttc aatgttggta tcaacaccag gtttaacttt gaacttatcg
     6481 gcactgacgg ttactgattt tgaacttttg ctttgccacg gaacggtctg cgttgtcggg
     6541 aagatgcgtg atctgatcct tcaactcagc aaaagttccc aatacgcaaa ccgcctctcc
     6601 ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg
     6661 gcagtgagcg caacgcaatt aatgtgagtt agctcactca ttaggcaccc caggctttac
     6721 actttatgct tccggctcgt atgttgtgtg gaattgtgag cggataacaa tttcacacag
     6781 gaaacagcta tgaccatgat tacgccaagc ttgcatgcct gcaggtcgac tctagaggat
     6841 ccccgggtac cgagctcgaa ttagcttgca tgcctgcagg tcaattccct ggcattatgc
     6901 ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc
     6961 tattaccatg
//
