LOCUS       sequence-131821-        8431 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express FLPe under EF-1 promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8431)
  TITLE     lenti vector
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8431)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pCDH-EF1-FLPe
FEATURES             Location/Qualifiers
     source          1..8431
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             234..414
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    458..583
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1076..1309
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1493..1537
                     /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    1804..1921
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2013..3191
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          2244..3182
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     3139..3159
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             join(3207..3212,3213..3233,3234..4505)
                     /codon_start=1
                     /product="nuclear-targeted site-specific recombinase"
                     /label=FLPe
                     /note="FLPe is a fusion between the SV40 nuclear 
                     localization signal and a thermostable version of the 
                     Saccharomyces cerevisiae site-specific recombinase FLP. 
                     Recombination occurs at FRT sequences (Buchholz et al., 
                     1998)."
                     /translation="MAPKKKRKVMSQFDILCKTPPKVLVRQFVERFERPSGEKIASCAA
                     ELTYLCWMITHNGTAIKRATFMSYNTIISNSLSFDIVNKSLQFKYKTQKATILEASLKK
                     LIPAWEFTIIPYNGQKHQSDITDIVSSLQLQFESSEEADKGNSHSKKMLKALLSEGESI
                     WEITEKILNSFEYTSRFTKTKTLYQFLFLATFINCGRFSDIKNVDPKSFKLVQNKYLGV
                     IIQCLVTETKTSVSRHIYFFSARGRIDPLVYLDEFLRNSEPVLKRVNRTGNSSSNKQEY
                     QLLKDNLVRSYNKALKKNAPYPIFAIKNGPKSHIGRHLMTSFLSMKGLTELTNVVGNWS
                     DKRASAVARTTYTHQITAIPDHYFALVSRYYAYDPISKEMIALKDETNPIEEWQHIEQL
                     KGSAEGSIRYPAWNGIISQEVLDYLSSYINRRI"
     misc_feature    4526..5114
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4579..4599)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4997..5008)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             5188..5421
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    5493..5614
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5530..5549)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5584..5603
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      5633..5768
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(5806..5822)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5806..5822)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5819..5841)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5830..5846
                     /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(5854..5860,5861..5878,5879..5884))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5899..5920
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6037..6054)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6208..6796)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(join(6967..7758,7759..7827))
                     /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     7590..7609
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7828..7932)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8000..8018
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(8056..8078)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     8178..8197
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     8391..8413
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     8405..8422
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     8406..8422
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 acgcgtgtag tcttatgcaa tactcttgta gtcttgcaac atggtaacga tgagttagca
       61 acatgcctta caaggagaga aaaagcaccg tgcatgccga ttggtggaag taaggtggta
      121 cgatcgtgcc ttattaggaa ggcaacagac gggtctgaca tggattggac gaaccactga
      181 attgccgcat tgcagagata ttgtatttaa gtgcctagct cgatacaata aacgggtctc
      241 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta
      301 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact
      361 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtggcg
      421 cccgaacagg gacctgaaag cgaaagggaa accagagctc tctcgacgca ggactcggct
      481 tgctgaagcg cgcacggcaa gaggcgaggg gcggcgactg gtgagtacgc caaaaatttt
      541 gactagcgga ggctagaagg agagagatgg gtgcgagagc gtcagtatta agcgggggag
      601 aattagatcg cgatgggaaa aaattcggtt aaggccaggg ggaaagaaaa aatataaatt
      661 aaaacatata gtatgggcaa gcagggagct agaacgattc gcagttaatc ctggcctgtt
      721 agaaacatca gaaggctgta gacaaatact gggacagcta caaccatccc ttcagacagg
      781 atcagaagaa cttagatcat tatataatac agtagcaacc ctctattgtg tgcatcaaag
      841 gatagagata aaagacacca aggaagcttt agacaagata gaggaagagc aaaacaaaag
      901 taagaccacc gcacagcaag cggccactga tcttcagacc tggaggagga gatatgaggg
      961 acaattggag aagtgaatta tataaatata aagtagtaaa aattgaacca ttaggagtag
     1021 cacccaccaa ggcaaagaga agagtggtgc agagagaaaa aagagcagtg ggaataggag
     1081 ctttgttcct tgggttcttg ggagcagcag gaagcactat gggcgcagcc tcaatgacgc
     1141 tgacggtaca ggccagacaa ttattgtctg gtatagtgca gcagcagaac aatttgctga
     1201 gggctattga ggcgcaacag catctgttgc aactcacagt ctggggcatc aagcagctcc
     1261 aggcaagaat cctggctgtg gaaagatacc taaaggatca acagctcctg gggatttggg
     1321 gttgctctgg aaaactcatt tgcaccactg ctgtgccttg gaatgctagt tggagtaata
     1381 aatctctgga acagattgga atcacacgac ctggatggag tgggacagag aaattaacaa
     1441 ttacacaagc ttaatacact ccttaattga agaatcgcaa aaccagcaag aaaagaatga
     1501 acaagaatta ttggaattag ataaatgggc aagtttgtgg aattggttta acataacaaa
     1561 ttggctgtgg tatataaaat tattcataat gatagtagga ggcttggtag gtttaagaat
     1621 agtttttgct gtactttcta tagtgaatag agttaggcag ggatattcac cattatcgtt
     1681 tcagacccac ctcccaaccc cgaggggacc cgacaggccc gaaggaatag aagaagaagg
     1741 tggagagaga gacagagaca gatccattcg attagtgaac ggatctcgac ggtatcggtt
     1801 aacttttaaa agaaaagggg ggattggggg gtacagtgca ggggaaagaa tagtagacat
     1861 aatagcaaca gacatacaaa ctaaagaatt acaaaaacaa attacaaaaa ttcaaaattt
     1921 tatcgatagc tttgcaaaga tggataaagt tttaaacaga gaggaatctt tgcagctaat
     1981 ggaccttcta ggtcttgaaa ggagtgggaa ttggctccgg tgcccgtcag tgggcagagc
     2041 gcacatcgcc cacagtcccc gagaagttgg ggggaggggt cggcaattga accggtgcct
     2101 agagaaggtg gcgcggggta aactgggaaa gtgatgtcgt gtactggctc cgcctttttc
     2161 ccgagggtgg gggagaaccg tatataagtg cagtagtcgc cgtgaacgtt ctttttcgca
     2221 acgggtttgc cgccagaaca caggtaagtg ccgtgtgtgg ttcccgcggg cctggcctct
     2281 ttacgggtta tggcccttgc gtgccttgaa ttacttccac ctggctgcag tacgtgattc
     2341 ttgatcccga gcttcgggtt ggaagtgggt gggagagttc gaggccttgc gcttaaggag
     2401 ccccttcgcc tcgtgcttga gttgaggcct ggcctgggcg ctggggccgc cgcgtgcgaa
     2461 tctggtggca ccttcgcgcc tgtctcgctg ctttcgataa gtctctagcc atttaaaatt
     2521 tttgatgacc tgctgcgacg ctttttttct ggcaagatag tcttgtaaat gcgggccaag
     2581 atctgcacac tggtatttcg gtttttgggg ccgcgggcgg cgacggggcc cgtgcgtccc
     2641 agcgcacatg ttcggcgagg cggggcctgc gagcgcggcc accgagaatc ggacgggggt
     2701 agtctcaagc tggccggcct gctctggtgc ctggcctcgc gccgccgtgt atcgccccgc
     2761 cctgggcggc aaggctggcc cggtcggcac cagttgcgtg agcggaaaga tggccgcttc
     2821 ccggccctgc tgcagggagc tcaaaatgga ggacgcggcg ctcgggagag cgggcgggtg
     2881 agtcacccac acaaaggaaa agggcctttc cgtcctcagc cgtcgcttca tgtgactcca
     2941 cggagtaccg ggcgccgtcc aggcacctcg attagttctc gagcttttgg agtacgtcgt
     3001 ctttaggttg gggggagggg ttttatgcga tggagtttcc ccacactgag tgggtggaga
     3061 ctgaagttag gccagcttgg cacttgatgt aattctcctt ggaatttgcc ctttttgagt
     3121 ttggatcttg gttcattctc aagcctcaga cagtggttca aagttttttt cttccatttc
     3181 aggtgtcgtg aggaattgga tccaccatgg ctcccaagaa gaagaggaag gtgatgagtc
     3241 aatttgatat attatgtaaa acaccaccta aggtcctggt tcgtcagttt gtggaaaggt
     3301 ttgaaagacc ttcaggggaa aaaatagcat catgtgctgc tgaactaacc tatttatgtt
     3361 ggatgattac tcataacgga acagcaatca agagagccac attcatgagc tataatacta
     3421 tcataagcaa ttcgctgagt ttcgatattg tcaacaaatc actccagttt aaatacaaga
     3481 cgcaaaaagc aacaattctg gaagcctcat taaagaaatt aattcctgct tgggaattta
     3541 caattattcc ttacaatgga caaaaacatc aatctgatat cactgatatt gtaagtagtt
     3601 tgcaattaca gttcgaatca tcggaagaag cagataaggg aaatagccac agtaaaaaaa
     3661 tgcttaaagc acttctaagt gagggtgaaa gcatctggga gatcactgag aaaatactaa
     3721 attcgtttga gtatacctcg agatttacaa aaacaaaaac tttataccaa ttcctcttcc
     3781 tagctacttt catcaattgt ggaagattca gcgatattaa gaacgttgat ccgaaatcat
     3841 ttaaattagt ccaaaataag tatctgggag taataatcca gtgtttagtg acagagacaa
     3901 agacaagcgt tagtaggcac atatacttct ttagcgcaag gggtaggatc gatccacttg
     3961 tatatttgga tgaatttttg aggaattctg aaccagtcct aaaacgagta aataggaccg
     4021 gcaattcttc aagcaacaaa caggaatacc aattattaaa agataactta gtcagatcgt
     4081 acaacaaggc tttgaagaaa aatgcgcctt atccaatctt tgctataaag aatggcccaa
     4141 aatctcacat tggaagacat ttgatgacct catttctgtc aatgaagggc ctaacggagt
     4201 tgactaatgt tgtgggaaat tggagcgata agcgtgcttc tgccgtggcc aggacaacgt
     4261 atactcatca gataacagca atacctgatc actacttcgc actagtttct cggtactatg
     4321 catatgatcc aatatcaaag gaaatgatag cattgaagga tgagactaat ccaattgagg
     4381 agtggcagca tatagaacag ctaaagggta gtgctgaagg aagcatacga taccccgcat
     4441 ggaatgggat aatatcacag gaggtactag actacctttc atcctacata aatagacgca
     4501 tataagcggc cgcgaattcg tcgacaatca acctctggat tacaaaattt gtgaaagatt
     4561 gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc
     4621 tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg
     4681 gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac
     4741 tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc
     4801 cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc
     4861 ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa
     4921 atcatcgtcc tttccttggc tgctcgcctg tgttgccacc tggattctgc gcgggacgtc
     4981 cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc
     5041 ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg
     5101 ggccgcctcc ccgcctggta cctttaagac caatgactta caaggcagct gtagatctta
     5161 gccacttttt aaaagaaaag gggggactgg aagggctaat tcactcccaa cgaaaataag
     5221 atctgctttt tgcttgtact gggtctctct ggttagacca gatctgagcc tgggagctct
     5281 ctggctaact agggaaccca ctgcttaagc ctcaataaag cttgccttga gtgcttcaag
     5341 tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag atccctcaga cccttttagt
     5401 cagtgtggaa aatctctagc agtagtagtt catgtcatct tattattcag tatttataac
     5461 ttgcaaagaa atgaatatca gagagtgaga ggaacttgtt tattgcagct tataatggtt
     5521 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     5581 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctggctctag ctatcccgcc
     5641 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
     5701 tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt
     5761 tggaggccta gacttttgca gagacggccc aaattcgtaa tcatggtcat agctgtttcc
     5821 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
     5881 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     5941 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     6001 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     6061 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     6121 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     6181 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     6241 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     6301 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     6361 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     6421 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     6481 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     6541 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     6601 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg
     6661 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     6721 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     6781 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     6841 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     6901 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     6961 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     7021 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     7081 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     7141 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     7201 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     7261 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     7321 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     7381 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     7441 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     7501 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     7561 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     7621 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     7681 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     7741 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     7801 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     7861 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     7921 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat
     7981 catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg
     8041 tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta
     8101 agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg
     8161 gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc atatgcggtg
     8221 tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggcgccatt cgccattcag
     8281 gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctggc
     8341 gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt cccagtcacg
     8401 acgttgtaaa acgacggcca gtgccaagct g
//
