LOCUS       sequence-190163-        7016 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor vector for 3' FLAG tag of human PBX2 .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7016)
  TITLE     Mendenhall CRISPR tagging plasmids
REFERENCE   2  (bases 1 to 7016)
  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_PBX2
FEATURES             Location/Qualifiers
     source          1..7016
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(731..1525)
                     /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(842..861)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     1452..1471
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     CDS             complement(1538..1594)
                     /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(1610..1633)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             complement(1637..1660)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             complement(1664..1687)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             complement(2813..3943)
                     /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(4019..4043)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        complement(4040..4242)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(4069..4089)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     complement(4419..4435)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4419..4435)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4432..4454)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4443..4459
                     /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(4467..4473,4474..4491,4492..4497))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4512..4533
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4833..4850)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5004..5592)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5084..5103)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(5961..5980)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             5972..6787
                     /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(6045..6064)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     6991..7013
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(7006..7016,1..6)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 gccagtgaac gacggccagt gaattggagg ctacagtcag tggagaggac tttccaaggt
       61 tcaaccccaa agcccagtca ataattccct aaagtagcag aaactccctc cgaggtagat
      121 atctgagtca gacactctcg tccaccgagc gattctattg gtttaagatg agctgcgtat
      181 gaggtaagta agccgtccgg aggggcgggg gtggggatgc atgggggcgt ggcccatgtc
      241 ctctgtccag aagtcatgtc cccatttttg gcatctctga ttgggcaggg ctggcgtctc
      301 cacagattcc agagcataca agtggggtgg ggaagggaaa gtgggggagc ccaggagaga
      361 aacagaatag ttgcaagtgg gagtatgtgt gtgtgaggtg tgggagaggg agagagaaag
      421 acagaggaga aaaaggggtc tgagaaatag gtttctcggt atgtgtatgt ttctgtgtaa
      481 gaaagaaagc gagagaggaa aaagatggaa aaaagggaga gacagacccc acactcccct
      541 tagaggcccc attcttcctg ccatgtaatt agcaccccca gcacagagag tctcgttagg
      601 gaggggatga ccccattggc ccttctctgt cttgtgcttc tcctgtattg gggtttgtcc
      661 tctggaagcc tgcgtcctct tcaagtcgcc ttgtgagagc ccccacccct gtgaccctga
      721 gatgttcgaa tcagaagaac tcgtcaagaa ggcgatagaa ggcgatgcgc tgcgaatcgg
      781 gagcggcgat accgtaaagc acgaggaagc ggtcagccca ttcgccgcca agctcttcag
      841 caatatcacg ggtagccaac gctatgtcct gatagcggtc ggccacaccc agccggccac
      901 agtcgatgaa tccagaaaag cggccatttt ccaccatgat attcggcaag caggcatcgc
      961 catgggtcac gacgagatcc tcgccgtcgg gcatgcgcgc cttgagcctg gcgaacagtt
     1021 cggctggcgc gagcccctga tgctcttcgt ccagatcatc ctgatcgaca agaccggctt
     1081 ccatccgagt acgtgctcgc tcgatgcgat gtttcgcttg gtggtcgaat gggcaggtag
     1141 ccggatcaag cgtatgcagc cgccgcattg catcagccat gatggatact ttctcggcag
     1201 gagcaaggtg agatgacagg agatcctgcc ccggcacttc gcccaatagc agccagtccc
     1261 ttcccgcttc agtgacaacg tcgagcacag ctgcgcaagg aacgcccgtc gtggccagcc
     1321 acgatagccg cgctgcctcg tcctgcagtt cattcagggc accggacagg tcggtcttga
     1381 caaaaagaac cgggcgcccc tgcgctgaca gccggaacac ggcggcatca gagcagccga
     1441 ttgtctgttg tgcccagtca tagccgaata gcctctccac ccaagcggcc ggagaacctg
     1501 cgtgcaatcc atcttgttca atcatgcgaa acgatccagg tccagggttc tcctccacgt
     1561 ctccagcctg cttcagcagg ctgaagttag tagctccgct tcctgaaacc ttatcgtcgt
     1621 catccttgta atctccctta tcgtcgtcat ccttgtaatc gcccttatcg tcgtcatcct
     1681 tgtaatctcc agaaccacct ccaccggaac ctcctccgct cccgttggag gtatcagagt
     1741 gaacactccc tggtccctcc gttggggatg tcactgaaga gggggtcaca gcctcttgcc
     1801 agctgccatt tgcctgaaag gagagacaga gtacagaaaa cagagaaagc cctgggaacc
     1861 ctgtgtgggc acaacattac tagggaaaat gcccctctgt cctgtgagaa ctggacagag
     1921 aggagcttca ggatccactc accctcattt cccgtgggct gtacatctgg cctcccccga
     1981 ggttatcccc atagccccct ggccccatcg agtgtcggag tgattccacc tgcaggcagc
     2041 agaggaaggt atgacagtga agagaagcct cagaggaaag aggtcttgta tcctaaagta
     2101 gaggaaatgg agttggggaa agccctattc gagaggagat gggcatctga cctgggaagc
     2161 agaataggaa tctccgttga gcccaggcat ccccagaaac atgtctccag atcctgagag
     2221 attgaaagag ccgccagagc cttgtggggg cagagaggga agagtgtaat agagcccgtg
     2281 atggtagagg atgaaccaca actctcaact cttgtgggga catgctacta tactccaatt
     2341 atccacaaaa taacattcca acacacagaa agagcaggct gttccttggc cacccgtggg
     2401 aagaaaggca gaactaagat cactggaatg gcctctgtcc cctgacatct ccagcctatc
     2461 tcagctcggt ccctctcacc ccaaaaggcc ccctctctgc tatgatcctg cctagatagg
     2521 aagagctggg ctgcaggtcg gggagaggag tgttagtaac cgcgacgctg gtgggggtcg
     2581 gcttgttaag aggggcgctg ctaacgctgc aagagtgggt tgtcagcgtg gggccggtgc
     2641 tactggaatc gataccggca tgattgacag cctgggcgag gatgtcacct gatggtgata
     2701 agaatacacg ggagacttag tacggtttca caggcgtgac acgtttattg agtaggatta
     2761 cagagtataa catagagtat aatatagagt atacaatagt gacgtgggat cctcagttag
     2821 cctcccccat ctcccgggca aacgtgcgcg ccaggtcgca gatcgtcggt atggagccgg
     2881 gggtggtgac gtgggtctgg accatcccgg aggtaagttg cagcagggcg tcccggcagc
     2941 cggcgggcga ttggtcgtaa tccaggataa agacgtgcat gggacggagg cgtttggcca
     3001 agacgtccaa ggcccaggca aacacgttgt acaggtcgcc gttgggggcc agcaactcgg
     3061 gggcccgaaa cagggtaaat aacgtgtccc cgatatgggg tcgtgggccc gcgttgctct
     3121 ggggctcggc accctggggc ggcacggccg tccccgaaag ctgtccccaa tcctcccgcc
     3181 acgacccgcc gccctgcaga taccgcaccg tattggcaag cagcccgtaa acgcggcgaa
     3241 tcgcggccag catagccagg tcaagccgct cgccggggcg ctggcgtttg gccaggcggt
     3301 cgatgtgtct gtcctccgga agggccccca acacgatgtt tgtgccgggc aaggtcggcg
     3361 ggatgagggc cacgaacgcc agcacggcct ggggggtcat gctgcccata aggtatcgcg
     3421 cggccgggta gcacaggagg gcggcgatgg gatggcggtc gaagatgagg gtgagggccg
     3481 ggggcggggc atgtgagctc ccagcctccc ccccgatatg aggagccaga acggcgtcgg
     3541 tcacggcata aggcatgccc attgttatct gggcgcttgt cattaccacc gccgcgtccc
     3601 cggccgatat ctcaccctgg tcgaggcggt gttgtgtggt gtagatgttc gcgattgtct
     3661 cggaagcccc cagcacctgc cagtaagtca tcggctcggg tacgtagacg atatcgtcgc
     3721 gcgaacccag ggccaccagc agttgcgtgg tggtggtttt ccccatcccg tgaggaccgt
     3781 ctatataaac ccgcagtagc gtgggcattt tctgctccag gcggacttcc gtggcttctt
     3841 gctgccggcg agggcgcaac gccgtacgtc ggttgctatg gccgcgagaa cgcgcagcct
     3901 ggtcgaacgc agacgcgtgt tgatggcagg ggtacgaagc catggtcggg atccccaggc
     3961 tggatcggtc ccggtgtctc ctatggaggt caaaacagcg tggatggcgt ctccaggcga
     4021 tctgacggtt cactaaacga gctctgctta tatagacctc ccaccgtaca cgcctaccgc
     4081 ccatttgcgt caatggggcg gagttgttac gacattttgg aaagtcccgt tgattttggt
     4141 gccaaaacaa actcccattg acgtcaatgg ggtggagact tgaaatcccc gtgagtcaaa
     4201 ccgctatcca cgcccattga tgtactgcca aaaccgcatc accatggtaa tagcgatgac
     4261 taatacgtag atgtactgcc aagtaggaaa gtcccataag gtcatgtact gggcataatg
     4321 ccagggaatt gacctgcagg catgcaagct aattcgagct cggtacccgg ggatcctcta
     4381 gagtcgacct gcaggcatgc aagcttggcg taatcatggt catagctgtt tcctgtgtga
     4441 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
     4501 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
     4561 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     4621 ggtttgcgta ttgggaactt ttgctgagtt gaaggatcag atcacgcatc ttcccgacaa
     4681 cgcagaccgt tccgtggcaa agcaaaagtt caaaatcagt aaccgtcagt gccgataagt
     4741 tcaaagttaa acctggtgtt gataccaaca ttgaaacgtt gatcgaaaac gcgctgaaaa
     4801 acgctgctga atgtgcgagc ttcttccgct tcctcgctca ctgactcgct gcgctcggtc
     4861 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     4921 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     4981 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     5041 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     5101 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     5161 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     5221 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     5281 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     5341 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     5401 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     5461 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     5521 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     5581 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaactcc
     5641 gtcgagaggt ctgcctcgtg aagaaggtgt tgctgactca taccaggcct gaatcgcccc
     5701 atcatccagc cagaaagtga gggagccacg gttgatgaga gctttgttgt aggtggacca
     5761 gttggtgatt ttgaactttt gctttgccac ggaacggtct gcgttgtcgg gaagatgcgt
     5821 gatctgatcc ttcaactcag caaaagttcg atttattcaa caaagccacg ttgtgtctca
     5881 aaatctctga tgttacattg cacaagataa aaatatatca tcatgaacaa taaaactgtc
     5941 tgcttacata aacagtaata caaggggtgt tatgagccat attcaacggg aaacgtcttg
     6001 ctcgaagccg cgattaaatt ccaacatgga tgctgattta tatgggtata aatgggctcg
     6061 cgataatgtc gggcaatcag gtgcgacaat ctatcgattg tatgggaagc ccgatgcgcc
     6121 agagttgttt ctgaaacatg gcaaaggtag cgttgccaat gatgttacag atgagatggt
     6181 cagactaaac tggctgacgg aatttatgcc tcttccgacc atcaagcatt ttatccgtac
     6241 tcctgatgat gcatggttac tcaccactgc gatccccggg aaaacagcat tccaggtatt
     6301 agaagaatat cctgattcag gtgaaaatat tgttgatgcg ctggcagtgt tcctgcgccg
     6361 gttgcattcg attcctgttt gtaattgtcc ttttaacagc gatcgcgtat ttcgtctcgc
     6421 tcaggcgcaa tcacgaatga ataacggttt ggttgatgcg agtgattttg atgacgagcg
     6481 taatggctgg cctgttgaac aagtctggaa agaaatgcat aagcttttgc cattctcacc
     6541 ggattcagtc gtcactcatg gtgatttctc acttgataac cttatttttg acgaggggaa
     6601 attaataggt tgtattgatg ttggacgagt cggaatcgca gaccgatacc aggatcttgc
     6661 catcctatgg aactgcctcg gtgagttttc tccttcatta cagaaacggc tttttcaaaa
     6721 atatggtatt gataatcctg atatgaatat attgcagttt catttgatgc tcgatgagtt
     6781 tttctaatca gaattggtta attggttgta acactggcag agcattacgc tgacttgacg
     6841 ggacggcggc tttgttgaat aaatcgcatt cgccattcag gctgcgcaac tgttgggaag
     6901 ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa
     6961 ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacg
//
