LOCUS       sequence-132103-        7164 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor vector for 3' FLAG tag of human HHEX .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7164)
  TITLE     Mendenhall CRISPR tagging plasmids
REFERENCE   2  (bases 1 to 7164)
  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"
COMMENT     Sequence Label: pFETCh_HHEX
FEATURES             Location/Qualifiers
     source          1..7164
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             847..870
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             874..897
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             901..924
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             940..996
                     /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             1009..1803
                     /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(1063..1082)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1673..1692
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(2666..2682)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2675..2697)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             complement(2901..3716)
                     /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     3624..3643
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3708..3727
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      4096..4684
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4585..4604
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4838..4855
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5155..5176
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5191..5196,5197..5214,5215..5221)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5229..5245
                     /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     5234..5256
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5253..5269
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5253..5269
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        5446..5648
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5599..5619
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     5645..5669
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             5745..6875
                     /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 cagttggcag cattcttcca gatccacatt ttttttcttt gccatgattt tgtaacttgt
       61 taaacattta cacatgtaga tgtttataaa tttactccag aaaatgaatg aaaaaagatc
      121 cagtctaata attgaaactt tattcttaat tcaccagtta gcattcttat ttttaaaatg
      181 tctgaaagac tgttaattgg caaggtttaa atatttaaag aatgaagtta aggtgtatct
      241 gtagtgatct gattttcttt cccccttaaa cttttgagaa tattgaaacc atgggtctgg
      301 gctggtgtta agtaaccaga atttgtgttc aattggtata atgtatctgc ccttatgttc
      361 atgactggtt ctcatctttt aagcatataa tctttgatat gacagaaact tttcttccaa
      421 agtgagcaaa aagcacaagt atgtatctcc tgaataagat tataaactga aaacactcag
      481 taaactaaaa attaaaagat ggaataggat gaagagacat tttgatttat tcctctcacc
      541 tatccatatt ttatgatcct tccaatctcc attttcattt cctttattca ggagaaccct
      601 caaagcaata aaaaagaaga actggaaagt ttggacagtt cctgtgatca gaggcaagat
      661 ttgcccagtg aacagaataa aggtgcttct ttggatagct ctcaatgttc gccctcccct
      721 gcctcccagg aagaccttga atcagagatt tcagaggatt ctgatcagga agtggacatt
      781 gagggcgata aaagctattt taatgctgga gggagcggag gaggttccgg tggaggtggt
      841 tctggagatt acaaggatga cgacgataag ggcgattaca aggatgacga cgataaggga
      901 gattacaagg atgacgacga taaggtttca ggaagcggag ctactaactt cagcctgctg
      961 aagcaggctg gagacgtgga ggagaaccct ggacctggat cgtttcgcat gattgaacaa
     1021 gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg
     1081 gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc
     1141 ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca
     1201 gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc
     1261 actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca
     1321 tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat
     1381 acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca
     1441 cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg
     1501 ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc
     1561 gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct
     1621 ggattcatcg actgtggccg gctgggtgtg gccgaccgct atcaggacat agcgttggct
     1681 acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac
     1741 ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc
     1801 tgattcgaac atccattggc atgttcagaa aactggattt aggaataatg ttttgctaca
     1861 gaaaatcttc atagaagaac tggaaggcta tataagaaag ggaatcaatt ctctggtatt
     1921 ctggaaacct aaaaatattt ggtgcactgc tcaattaaca aacctacatg gagaccttaa
     1981 ttttgactta acaaatagtt tatgtactgc tcttaggttg ttttgataaa gtgacattat
     2041 agtgattaaa ttcttccccc tttaaaaaaa cagttagtgg ttttcactat ttataaaaaa
     2101 ttaattttga actttttgtt aaatttttaa gttatagctt taaaggtttt aataggacct
     2161 tcttgaacga cttttctgta atctgtttat ctcccactta atggaaagac aaaggggtac
     2221 cccaaatcca gaggtgccta catttcatgc agccttggag tattttaaaa ggaaaacatt
     2281 ctttactttt atatgacatt cttatactgc tgtctcaaat ccaaaaacat ttcagagctc
     2341 ttgtctcaga gatgtgtgtt ctttttgtca gagatatggt tgatgagaat cttaaatgct
     2401 tgttttgcac tatcacttag tacctgtttg accaaggtgt taaggggata gtacctccca
     2461 attcaagcag agaaactgac ctgactaaag ttaatcgcag atgaactaga agtcacaggt
     2521 taattaaatg taagtagatt gtagatactg ttttatatca aacaatgttt ataatgtgta
     2581 tatagaattg ttcactgtaa aaaaaatggc caacttggaa agtcctctcc actgactgta
     2641 gcctccaatt cactggccgt cgttcactgg ccgtcgtttt acaacgtcgt gactgggaaa
     2701 accctggcgt tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta
     2761 atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat
     2821 gcgatttatt caacaaagcc gccgtcccgt caagtcagcg taatgctctg ccagtgttac
     2881 aaccaattaa ccaattctga ttagaaaaac tcatcgagca tcaaatgaaa ctgcaattta
     2941 ttcatatcag gattatcaat accatatttt tgaaaaagcc gtttctgtaa tgaaggagaa
     3001 aactcaccga ggcagttcca taggatggca agatcctggt atcggtctgc gattccgact
     3061 cgtccaacat caatacaacc tattaatttc ccctcgtcaa aaataaggtt atcaagtgag
     3121 aaatcaccat gagtgacgac tgaatccggt gagaatggca aaagcttatg catttctttc
     3181 cagacttgtt caacaggcca gccattacgc tcgtcatcaa aatcactcgc atcaaccaaa
     3241 ccgttattca ttcgtgattg cgcctgagcg agacgaaata cgcgatcgct gttaaaagga
     3301 caattacaaa caggaatcga atgcaaccgg cgcaggaaca ctgccagcgc atcaacaata
     3361 ttttcacctg aatcaggata ttcttctaat acctggaatg ctgttttccc ggggatcgca
     3421 gtggtgagta accatgcatc atcaggagta cggataaaat gcttgatggt cggaagaggc
     3481 ataaattccg tcagccagtt tagtctgacc atctcatctg taacatcatt ggcaacgcta
     3541 cctttgccat gtttcagaaa caactctggc gcatcgggct tcccatacaa tcgatagatt
     3601 gtcgcacctg attgcccgac attatcgcga gcccatttat acccatataa atcagcatcc
     3661 atgttggaat ttaatcgcgg cttcgagcaa gacgtttccc gttgaatatg gctcataaca
     3721 ccccttgtat tactgtttat gtaagcagac agttttattg ttcatgatga tatattttta
     3781 tcttgtgcaa tgtaacatca gagattttga gacacaacgt ggctttgttg aataaatcga
     3841 acttttgctg agttgaagga tcagatcacg catcttcccg acaacgcaga ccgttccgtg
     3901 gcaaagcaaa agttcaaaat caccaactgg tccacctaca acaaagctct catcaaccgt
     3961 ggctccctca ctttctggct ggatgatggg gcgattcagg cctggtatga gtcagcaaca
     4021 ccttcttcac gaggcagacc tctcgacgga gttccactga gcgtcagacc ccgtagaaaa
     4081 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     4141 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     4201 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     4261 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     4321 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     4381 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     4441 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     4501 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     4561 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     4621 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     4681 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     4741 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
     4801 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
     4861 ggaagaagct cgcacattca gcagcgtttt tcagcgcgtt ttcgatcaac gtttcaatgt
     4921 tggtatcaac accaggttta actttgaact tatcggcact gacggttact gattttgaac
     4981 ttttgctttg ccacggaacg gtctgcgttg tcgggaagat gcgtgatctg atccttcaac
     5041 tcagcaaaag ttcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg
     5101 cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt
     5161 gagttagctc actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt
     5221 gtgtggaatt gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgc
     5281 caagcttgca tgcctgcagg tcgactctag aggatccccg ggtaccgagc tcgaattagc
     5341 ttgcatgcct gcaggtcaat tccctggcat tatgcccagt acatgacctt atgggacttt
     5401 cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtgat gcggttttgg
     5461 cagtacatca atgggcgtgg atagcggttt gactcacggg gatttcaagt ctccacccca
     5521 ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta
     5581 acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa
     5641 gcagagctcg tttagtgaac cgtcagatcg cctggagacg ccatccacgc tgttttgacc
     5701 tccataggag acaccgggac cgatccagcc tggggatccc gaccatggct tcgtacccct
     5761 gccatcaaca cgcgtctgcg ttcgaccagg ctgcgcgttc tcgcggccat agcaaccgac
     5821 gtacggcgtt gcgccctcgc cggcagcaag aagccacgga agtccgcctg gagcagaaaa
     5881 tgcccacgct actgcgggtt tatatagacg gtcctcacgg gatggggaaa accaccacca
     5941 cgcaactgct ggtggccctg ggttcgcgcg acgatatcgt ctacgtaccc gagccgatga
     6001 cttactggca ggtgctgggg gcttccgaga caatcgcgaa catctacacc acacaacacc
     6061 gcctcgacca gggtgagata tcggccgggg acgcggcggt ggtaatgaca agcgcccaga
     6121 taacaatggg catgccttat gccgtgaccg acgccgttct ggctcctcat atcggggggg
     6181 aggctgggag ctcacatgcc ccgcccccgg ccctcaccct catcttcgac cgccatccca
     6241 tcgccgccct cctgtgctac ccggccgcgc gataccttat gggcagcatg accccccagg
     6301 ccgtgctggc gttcgtggcc ctcatcccgc cgaccttgcc cggcacaaac atcgtgttgg
     6361 gggcccttcc ggaggacaga cacatcgacc gcctggccaa acgccagcgc cccggcgagc
     6421 ggcttgacct ggctatgctg gccgcgattc gccgcgttta cgggctgctt gccaatacgg
     6481 tgcggtatct gcagggcggc gggtcgtggc gggaggattg gggacagctt tcggggacgg
     6541 ccgtgccgcc ccagggtgcc gagccccaga gcaacgcggg cccacgaccc catatcgggg
     6601 acacgttatt taccctgttt cgggcccccg agttgctggc ccccaacggc gacctgtaca
     6661 acgtgtttgc ctgggccttg gacgtcttgg ccaaacgcct ccgtcccatg cacgtcttta
     6721 tcctggatta cgaccaatcg cccgccggct gccgggacgc cctgctgcaa cttacctccg
     6781 ggatggtcca gacccacgtc accacccccg gctccatacc gacgatctgc gacctggcgc
     6841 gcacgtttgc ccgggagatg ggggaggcta actgaggatc ccacgtcact attgtatact
     6901 ctatattata ctctatgtta tactctgtaa tcctactcaa taaacgtgtc acgcctgtga
     6961 aaccgtacta agtctcccgt gtattcttat caccatcagg tgacatcctc gcccaggctg
     7021 tcaatcatgc cggtatcgat tccagtagca ccggccccac gctgacaacc cactcttgca
     7081 gcgttagcag cgcccctctt aacaagccga cccccaccag cgtcgcggtt actaacactc
     7141 ctctccccga cctgcagccc agct
//
