LOCUS       sequence-375060-        7455 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral Flp Recombinase driven by the EFS promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7455)
  AUTHORS   Orstad G, Fort G, Parnell TJ, Jones A, Stubben C, Lohman B, Gillis 
            KL, Orellana W, Tariq R, Klingbeil O, Kaestner K, Vakoc CR, Spike 
            BT, Snyder EL
  TITLE     FoxA1 and FoxA2 control growth and cellular identity in 
            NKX2-1-positive lung adenocarcinoma.
  JOURNAL   Dev Cell. 2022 Aug 8;57(15):1866-1882.e10. doi: 
            10.1016/j.devcel.2022.06.017. Epub 2022 Jul 13.
  PUBMED    35835117
REFERENCE   2  (bases 1 to 7455)
  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"; doi: 
            "10.1016/j.devcel.2022.06.017"; journalName: "Dev Cell"; date: 
            "2022-08-8- 8"; volume: "57"; issue: "15"; pages: "1866-1882.e10"
COMMENT     Sequence Label: pCDH-EFS-FlpO
FEATURES             Location/Qualifiers
     source          1..7455
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(344..355)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             535..768
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    840..961
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(877..896)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     931..950
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      1001..1136
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(1169..1185)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1169..1185)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1182..1204)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1193..1209
                     /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(1217..1223,1224..1241,1242..1247))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1262..1283
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1400..1417)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1571..2159)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(join(2330..3121,3122..3190))
                     /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     2953..2972
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3191..3295)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3363..3381
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3419..3441)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3541..3560
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3754..3776
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3768..3785
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3769..3785
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        3800..4026
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             4027..4207
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    4254..4379
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    4872..5105
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             5290..5334
                     /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    5601..5717
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        5787..5998
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     CDS             join(6015..6020,6021..6041,6042..7313)
                     /codon_start=1
                     /product="nuclear-targeted site-specific recombinase"
                     /label=FLPo
                     /note="FLPo is a mouse codon-optimized fusion between the 
                     SV40 nuclear localization signal and a thermostable version
                     of the Saccharomyces cerevisiae site-specific recombinase 
                     FLP.  Recombination occurs at FRT sequences."
                     /translation="MAPKKKRKVMSQFDILCKTPPKVLVRQFVERFERPSGEKIASCAA
                     ELTYLCWMITHNGTAIKRATFMSYNTIISNSLSFDIVNKSLQFKYKTQKATILEASLKK
                     LIPAWEFTIIPYNGQKHQSDITDIVSSLQLQFESSEEADKGNSHSKKMLKALLSEGESI
                     WEITEKILNSFEYTSRFTKTKTLYQFLFLATFINCGRFSDIKNVDPKSFKLVQNKYLGV
                     IIQCLVTETKTSVSRHIYFFSARGRIDPLVYLDEFLRNSEPVLKRVNRTGNSSSNKQEY
                     QLLKDNLVRSYNKALKKNAPYPIFAIKNGPKSHIGRHLMTSFLSMKGLTELTNVVGNWS
                     DKRASAVARTTYTHQITAIPDHYFALVSRYYAYDPISKEMIALKDETNPIEEWQHIEQL
                     KGSAEGSIRYPAWNGIISQEVLDYLSSYINRRI"
     misc_feature    join(7328..7455,1..461)
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(7381..7401)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
ORIGIN
        1 cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt
       61 tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg
      121 cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac
      181 ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac
      241 tgacaattcc gtggtgttgt cggggaaatc atcgtccttt ccttggctgc tcgcctgtgt
      301 tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc
      361 ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg
      421 ccctcagacg agtcggatct ccctttgggc cgcctccccg cctggtacct ttaagaccaa
      481 tgacttacaa ggcagctgta gatcttagcc actttttaaa agaaaagggg ggactggaag
      541 ggctaattca ctcccaacga agataagatc tgctttttgc ttgtactggg tctctctggt
      601 tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc
      661 aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta
      721 actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt agtagttcat
      781 gtcatcttat tattcagtat ttataacttg caaagaaatg aatatcagag agtgagagga
      841 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
      901 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
      961 atcatgtctg gctctagcta tcccgcccct aactccgccc atcccgcccc taactccgcc
     1021 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga
     1081 ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctaga
     1141 cttttgcaga gaccaaattc gtaatcatgt catagctgtt tcctgtgtga aattgttatc
     1201 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
     1261 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     1321 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     1381 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     1441 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     1501 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     1561 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     1621 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     1681 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     1741 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     1801 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     1861 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     1921 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     1981 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     2041 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     2101 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     2161 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     2221 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     2281 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     2341 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     2401 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     2461 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     2521 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     2581 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     2641 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     2701 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     2761 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     2821 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     2881 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     2941 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     3001 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     3061 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     3121 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     3181 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     3241 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     3301 ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata
     3361 aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc
     3421 tctgacacat gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca
     3481 gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg
     3541 cggcatcaga gcagattgta ctgagagtgc accatatgcg gtgtgaaata ccgcacagat
     3601 gcgtaaggag aaaataccgc atcaggcgcc attcgccatt caggctgcgc aactgttggg
     3661 aagggcgatc ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg
     3721 caaggcgatt aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg
     3781 ccagtgccaa gctgacgcgt gtagtcttat gcaatactct tgtagtcttg caacatggta
     3841 acgatgagtt agcaacatgc cttacaagga gagaaaaagc accgtgcatg ccgattggtg
     3901 gaagtaaggt ggtacgatcg tgccttatta ggaaggcaac agacgggtct gacatggatt
     3961 ggacgaacca ctgaattgcc gcattgcaga gatattgtat ttaagtgcct agctcgatac
     4021 ataaacgggt ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg
     4081 aacccactgc ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt
     4141 ctgttgtgtg actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc
     4201 tctagcagtg gcgcccgaac agggacttga aagcgaaagg gaaaccagag gagctctctc
     4261 gacgcaggac tcggcttgct gaagcgcgca cggcaagagg cgaggggcgg cgactggtga
     4321 gtacgccaaa aattttgact agcggaggct agaaggagag agatgggtgc gagagcgtca
     4381 gtattaagcg ggggagaatt agatcgcgat gggaaaaaat tcggttaagg ccagggggaa
     4441 agaaaaaata taaattaaaa catatagtat gggcaagcag ggagctagaa cgattcgcag
     4501 ttaatcctgg cctgttagaa acatcagaag gctgtagaca aatactggga cagctacaac
     4561 catcccttca gacaggatca gaagaactta gatcattata taatacagta gcaaccctct
     4621 attgtgtgca tcaaaggata gagataaaag acaccaagga agctttagac aagatagagg
     4681 aagagcaaaa caaaagtaag accaccgcac agcaagcggc cactgatctt cagacctgga
     4741 ggaggagata tgagggacaa ttggagaagt gaattatata aatataaagt agtaaaaatt
     4801 gaaccattag gagtagcacc caccaaggca aagagaagag tggtgcagag agaaaaaaga
     4861 gcagtgggaa taggagcttt gttccttggg ttcttgggag cagcaggaag cactatgggc
     4921 gcagcgtcaa tgacgctgac ggtacaggcc agacaattat tgtctggtat agtgcagcag
     4981 cagaacaatt tgctgagggc tattgaggcg caacagcatc tgttgcaact cacagtctgg
     5041 ggcatcaagc agctccaggc aagaatcctg gctgtggaaa gatacctaaa ggatcaacag
     5101 ctcctgggga tttggggttg ctctggaaaa ctcatttgca ccactgctgt gccttggaat
     5161 gctagttgga gtaataaatc tctggaacag atttggaatc acacgacctg gatggagtgg
     5221 gacagagaaa ttaacaatta cacaagctta atacactcct taattgaaga atcgcaaaac
     5281 cagcaagaaa agaatgaaca agaattattg gaattagata aatgggcaag tttgtggaat
     5341 tggtttaaca taacaaattg gctgtggtat ataaaattat tcataatgat agtaggaggc
     5401 ttggtaggtt taagaatagt ttttgctgta ctttctatag tgaatagagt taggcaggga
     5461 tattcaccat tatcgtttca gacccacctc ccaaccccga ggggacccga caggcccgaa
     5521 ggaatagaag aagaaggtgg agagagagac agagacagat ccattcgatt agtgaacgga
     5581 tctcgacggt atcggttaac ttttaaaaga aaagggggga ttggggggta cagtgcaggg
     5641 gaaagaatag tagacataat agcaacagac atacaaacta aagaattaca aaaacaaatt
     5701 acaaaattca aaattttatc gatactagta ttatgcccag tacatgacct tatgggactt
     5761 tcctacttgg cagtacatct actagtgggc agagcgcaca tcgcccacag tccccgagaa
     5821 gttgggggga ggggtcggca attgaacggg tgcctagaga aggtggcgcg gggtaaactg
     5881 ggaaagtgat gtcgtgtact ggctccgcct ttttcccgag ggtgggggag aaccgtatat
     5941 aagtgcagta gtcgccgtga acgttctttt tcgcaacggg tttgccgcca gaacacagct
     6001 gcagtctaga attcatggct cctaagaaga agaggaaggt gatgagccag ttcgacatcc
     6061 tgtgcaagac cccccccaag gtgctggtgc ggcagttcgt ggagagattc gagaggccca
     6121 gcggcgagaa gatcgccagc tgtgccgccg agctgaccta cctgtgctgg atgatcaccc
     6181 acaacggcac cgccatcaag agggccacct tcatgagcta caacaccatc atcagcaaca
     6241 gcctgagctt cgacatcgtg aacaagagcc tgcagttcaa gtacaagacc cagaaggcca
     6301 ccatcctgga ggccagcctg aagaagctga tccccgcctg ggagttcacc atcatccctt
     6361 acaacggcca gaagcaccag agcgacatca ccgacatcgt gtccagcctg cagctgcagt
     6421 tcgagagcag cgaggaggcc gacaagggca acagccacag caagaagatg ctgaaggccc
     6481 tgctgtccga gggcgagagc atctgggaga tcaccgagaa gatcctgaac agcttcgagt
     6541 acaccagcag gttcaccaag accaagaccc tgtaccagtt cctgttcctg gccacattca
     6601 tcaactgcgg caggttcagc gacatcaaga acgtggaccc caagagcttc aagctggtgc
     6661 agaacaagta cctgggcgtg atcattcagt gcctggtgac cgagaccaag acaagcgtgt
     6721 ccaggcacat ctactttttc agcgccagag gcaggatcga ccccctggtg tacctggacg
     6781 agttcctgag gaacagcgag cccgtgctga agagagtgaa caggaccggc aacagcagca
     6841 gcaacaagca ggagtaccag ctgctgaagg acaacctggt gcgcagctac aacaaggccc
     6901 tgaagaagaa cgccccctac cccatcttcg ctatcaagaa cggccctaag agccacatcg
     6961 gcaggcacct gatgaccagc tttctgagca tgaagggcct gaccgagctg acaaacgtgg
     7021 tgggcaactg gagcgacaag agggcctccg ccgtggccag gaccacctac acccaccaga
     7081 tcaccgccat ccccgaccac tacttcgccc tggtgtccag gtactacgcc tacgacccca
     7141 tcagcaagga gatgatcgcc ctgaaggacg agaccaaccc catcgaggag tggcagcaca
     7201 tcgagcagct gaagggcagc gccgagggca gcatcagata ccccgcctgg aacggcatca
     7261 tcagccagga ggtgctggac tacctgagca gctacatcaa caggcggatc tgattaatta
     7321 agtcgacaat caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta
     7381 tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc
     7441 ttcccgtatg gcttt
//
