LOCUS       sequence-372924-        7714 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  lentiviral vector with Flag, HA tags.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7714)
  AUTHORS   Yousefzadeh MJ, Wyatt DW, Takata K, Mu Y, Hensley SC, Tomida J, 
            Bylund GO, Doublie S, Johansson E, Ramsden DA, McBride KM, Wood RD
  TITLE     Mechanism of suppression of chromosomal instability by DNA 
            polymerase POLQ.
  JOURNAL   PLoS Genet. 2014 Oct 2;10(10):e1004654. doi: 
            10.1371/journal.pgen.1004654. eCollection 2014 Oct.
  PUBMED    25275444
REFERENCE   2  (bases 1 to 7714)
  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: "PLoS 
            Genet."; date: "2014-10-2"; volume: "10(10):e1004654. doi"; pages: "
            10.1371/journal.pgen.1004654. eCollection 2014 Oct"
COMMENT     Sequence Label: pCDH-EF1-FHC
FEATURES             Location/Qualifiers
     source          1..7714
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     144..161
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     CDS             complement(339..365)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="This is a variant of the standard HA tag, which has 
                     sequence YPYDVPDYA. Some anti-HA antibodies may not 
                     recognize this variant."
                     /translation="YPYDVPDYA"
     CDS             complement(339..365)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             complement(381..404)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     LTR             476..744
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     promoter        complement(757..968)
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     misc_feature    1011..1127
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     CDS             complement(1394..1438)
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    1623..1856
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     misc_feature    2349..2474
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     LTR             2521..2701
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     promoter        complement(2702..2928)
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     primer_bind     complement(2943..2960)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2943..2959)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2952..2974)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(3168..3187)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3287..3309
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3347..3365)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3433..3537
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3538..3606,3607..4398)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(3756..3775)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4569..5157
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5311..5328
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5445..5466
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5481..5486,5487..5504,5505..5511)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5519..5535
                     /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     5524..5546
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5543..5559
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5543..5559
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     rep_origin      5592..5727
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    5767..5888
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5778..5797)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5832..5851
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     LTR             5960..6193
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     misc_feature    6267..6855
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             6373..6384
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     6782..6802
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6862..7461)
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(6945..6965)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     7442..7461
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     misc_feature    join(7470..7714,1..328)
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(7629..7648)
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
ORIGIN
        1 atcttataca cgtggctttt ggccgcagag gcacctgtcg ccaggtgggg ggttccgctg
       61 cctgcaaagg gtcgctacag acgttgtttg tcttcaagaa gcttccagag gaactgcttc
      121 cttcacgaca ttcaacagac cttgcattcc tttggcgaga ggggaaagac ccctaggaat
      181 gctcgtcaag aagacagggc caggtttccg ggccctcaca ttgccaaaag acggcaatat
      241 ggtggaaaat aacatataga caaacgcaca ccggccttat tccaagcggc ttcggccagt
      301 aacgttangg gggggggagg gagaggggcg cggccctagg cgtagtcggg cacgtcgtag
      361 gggtatcctc cagcggccga cttgtcatcg tcgtccttgt agtctcctcc agcggccgcg
      421 gatccgattt aaattcgaat tcgctagctc tagagtaggc gccggtcaca gcttggatct
      481 gtaacggcgc agaacagaaa acgaaacaaa gacgtagagt tgagcaagca gggtcaggca
      541 aagcgtggag agccggctga gtctaggtag gctccaaggg agcgccggac aaaggcccgg
      601 tctcgacctg agctttaaac ttacctagac ggcggacgca gttcaggagg caccacaggc
      661 gggaggcggc agaacgcgac tcaaccggcg tggatggcgg cctcaggtag ggcggcgggc
      721 gcgtgaagga gagatgcgag cccctcgaag cttcagctgt gttctggcgg caaacccgtt
      781 gcgaaaaaga acgttcacgg cgactactgc acttatatac ggttctcccc caccctcggg
      841 aaaaaggcgg agccagtaca cgacatcact ttcccagttt accccgcgcc accttctcta
      901 ggcacccgtt caattgccga cccctccccc caacttctcg gggactgtgg gcgatgtgcg
      961 ctctgcccac tgacgggcac cggagcgatc gcagatccac tagtatcgat aaaattttga
     1021 attttgtaat ttgtttttgt aattctttag tttgtatgtc tgttgctatt atgtctacta
     1081 ttctttcccc tgcactgtac cccccaatcc ccccttttct tttaaaagtt aaccgatacc
     1141 gtcgagatcc gttcactaat cgaatggatc tgtctctgtc tctctctcca ccttcttctt
     1201 ctattccttc gggcctgtcg ggtcccctcg gggttgggag gtgggtctga aacgataatg
     1261 gtgaatatcc ctgcctaact ctattcacta tagaaagtac agcaaaaact attcttaaac
     1321 ctaccaagcc tcctactatc attatgaata attttatata ccacagccaa tttgttatgt
     1381 taaaccaatt ccacaaactt gcccatttat ctaattccaa taattcttgt tcattctttt
     1441 cttgctggtt ttgcgattct tcaattaagg agtgtattaa gcttgtgtaa ttgttaattt
     1501 ctctgtccca ctccatccag gtcgtgtgat tccaaatctg ttccagagat ttattactcc
     1561 aactagcatt ccaaggcaca gcagtggtgc aaatgagttt tccagagcaa ccccaaatcc
     1621 ccaggagctg ttgatccttt aggtatcttt ccacagccag gattcttgcc tggagctgct
     1681 tgatgcccca gactgtgagt tgcaacagat gctgttgcgc ctcaatagcc ctcagcaaat
     1741 tgttctgctg ctgcactata ccagacaata attgtctggc ctgtaccgtc agcgtcattg
     1801 acgctgcgcc catagtgctt cctgctgctc ccaagaaccc aaggaacaaa gctcctattc
     1861 ccactgctct tttttctctc tgcaccactc ttctctttgc cttggtgggt gctactccta
     1921 atggttcaat ttttactact ttatatttat ataattcact tctccaattg tccctcatat
     1981 ctcctcctcc aggtctgaag atcagtggcc gcttgctgtg cggtggtctt acttttgttt
     2041 tgctcttcct ctatcttgtc taaagcttcc ttggtgtctt ttatctctat cctttgatgc
     2101 acacaataga gggttgctac tgtattatat aatgatctaa gttcttctga tcctgtctga
     2161 agggatggtt gtagctgtcc cagtatttgt ctacagcctt ctgatgtttc taacaggcca
     2221 ggattaactg cgaatcgttc tagctccctg cttgcccata ctatatgttt taatttatat
     2281 tttttctttc cccctggcct taaccgaatt ttttcccatc gcgatctaat tctcccccgc
     2341 ttaatactga cgctctcgca cccatctctc tccttctagc ctccgctagt caaaattttt
     2401 ggcgtactca ccagtcgccg cccctcgcct cttgccgtgc gcgcttcagc aagccgagtc
     2461 ctgcgtcgag agagctcctc tggtttccct ttcgctttca agtccctgtt cgggcgccac
     2521 tgctagagat tttccacact gactaaaagg gtctgaggga tctctagtta ccagagtcac
     2581 acaacagacg ggcacacact acttgaagca ctcaaggcaa gctttattga ggcttaagca
     2641 gtgggttccc tagttagcca gagagctccc aggctcagat ctggtctaac cagagagacc
     2701 cgtttatgta tcgagctagg cacttaaata caatatctct gcaatgcggc aattcagtgg
     2761 ttcgtccaat ccatgtcaga cccgtctgtt gccttcctaa taaggcacga tcgtaccacc
     2821 ttacttccac caatcggcat gcacggtgct ttttctctcc ttgtaaggca tgttgctaac
     2881 tcatcgttac catgttgcaa gactacaaga gtattgcata agactacacg cgtcagcttg
     2941 gcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat
     3001 cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat
     3061 cgcccttccc aacagttgcg cagcctgaat ggcgaatggc gcctgatgcg gtattttctc
     3121 cttacgcatc tgtgcggtat ttcacaccgc atatggtgca ctctcagtac aatctgctct
     3181 gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg
     3241 gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg
     3301 tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc
     3361 ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt
     3421 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     3481 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     3541 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     3601 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     3661 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     3721 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     3781 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     3841 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     3901 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     3961 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     4021 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     4081 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     4141 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     4201 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     4261 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     4321 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     4381 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     4441 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     4501 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     4561 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     4621 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     4681 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     4741 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     4801 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     4861 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     4921 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     4981 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     5041 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     5101 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     5161 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     5221 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
     5281 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag
     5341 cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac
     5401 gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc
     5461 actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt
     5521 gtgagcggat aacaatttca cacaggaaac agctatgaca tgattacgaa tttggtctct
     5581 gcaaaagtct aggcctccaa aaaagcctcc tcactacttc tggaatagct cagaggccga
     5641 ggcggcctcg gcctctgcat aaataaaaaa aattagtcag ccatggggcg gagaatgggc
     5701 ggaactgggc ggagttaggg gcgggatggg cggagttagg ggcgggatag ctagagccag
     5761 acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat
     5821 gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata
     5881 aacaagttcc tctcactctc tgatattcat ttctttgcaa gttataaata ctgaataata
     5941 agatgacatg aactactact gctagagatt ttccacactg actaaaaggg tctgagggat
     6001 ctctagttac cagagtcaca caacagacgg gcacacacta cttgaagcac tcaaggcaag
     6061 ctttattgag gcttaagcag tgggttccct agttagccag agagctccca ggctcagatc
     6121 tggtctaacc agagagaccc agtacaagca aaaagcagat cttatcttcg ttgggagtga
     6181 attagccctt ccagtccccc cttttctttt aaaaagtggc taagatctac agctgccttg
     6241 taagtcattg gtcttaaagg taccaggcgg ggaggcggcc caaagggaga tccgactcgt
     6301 ctgagggcga aggcgaagac gcggaagagg ccgcagagcc ggcagcaggc cgcgggaagg
     6361 aaggtccgct ggattgaggg ccgaagggac gtagcagaag gacgtcccgc gcagaatcca
     6421 ggtggcaaca caggcgagca gccaaggaaa ggacgatgat ttccccgaca acaccacgga
     6481 attgtcagtg cccaacagcc gagcccctgt ccagcagcgg gcaaggcagg cggcgatgag
     6541 ttccgccgtg gcaataggga gggggaaagc gaaagtcccg gaaaggagct gacaggtggt
     6601 ggcaatgccc caaccagtgg gggttgcgtc agcaaacaca gtgcacacca cgccacgttg
     6661 cctgacaacg ggccacaact cctcataaag agacagcaac caggatttat acaaggagga
     6721 gaaaatgaaa gccatacggg aagcaatagc atgatacaaa ggcattaaag cagcgtatcc
     6781 acatagcgta aaaggagcaa catagttaag aataccagtc aatctttcac aaattttgta
     6841 atccagaggt tgattgtcga cctaggcacc gggcttgcgg gtcatgcacc aggtgcgcgg
     6901 tccttcgggc acctcgacgt cggcggtgac ggtgaagccg agccgctcgt agaaggggag
     6961 gttgcggggc gcggaggtct ccaggaaggc gggcaccccg gcgcgctcgg ccgcctccac
     7021 tccggggagc acgacggcgc tgcccagacc cttgccctgg tggtcgggcg agacgccgac
     7081 ggtggccagg aaccacgcgg gctccttggg ccggtgcggc gccaggaggc cttccatctg
     7141 ttgctgcgcg gccagccggg aaccgctcaa ctcggccatg cgcgggccga tctcggcgaa
     7201 caccgccccc gcttcgacgc tctccggcgt ggtccagacc gccaccgcgg cgccgtcgtc
     7261 cgcgacccac accttgccga tgtcgagccc gacgcgcgtg aggaagagtt cttgcagctc
     7321 ggtgacccgc tcgatgtggc ggtccggatc gacggtgtgg cgcgtggcgg ggtagtcggc
     7381 gaacgcggcg gcgagggtgc gtacggccct ggggacgtcg tcgcgggtgg cgaggcgcac
     7441 cgtgggcttg tactcggtca ttccggaggt attatcgtgt ttttcaaagg aaaaccacgt
     7501 ccccgtggtt cggggggcct agacgttttt ttaacctcga ctaaacacat gtaaagcatg
     7561 tgcaccgagg ccccagatca gatcccatac aatggggtac cttctgggca tccttcagcc
     7621 ccttgttgaa tacgcttgag gagagccatt tgactctttc cacaactatc caactcacaa
     7681 cgtggcactg gggttgtgcc gcctttgcag gtgt
//
