LOCUS       sequence-106042-        7563 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cre-regulated lentiviral shRNA vector. Cre addition causes EGFP to 
            be recombined out of the construct, activating shRNA expression..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7563)
  AUTHORS   Ventura A, Meissner A, Dillon CP, McManus M, Sharp PA, Van Parijs L,
            Jaenisch R, Jacks T
  TITLE     Cre-lox-regulated conditional RNA interference from transgenes.
  JOURNAL   Proc Natl Acad Sci U S A. 2004 Jul 13. 101(28):10380-5.
  PUBMED    15240889
REFERENCE   2  (bases 1 to 7563)
  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: "Proc Natl 
            Acad Sci U S A. 2004 Jul 13. 101(28):10380-5."
COMMENT     Sequence Label: pSico
FEATURES             Location/Qualifiers
     source          1..7563
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             337..381
                     /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    679..796
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     primer_bind     1086..1106
                     /label=mU6-F
                     /note="Mouse U6 promoter, forward primer"
     protein_bind    complement(1124..1157)
                     /label=loxP511
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATAC) (Shaw et al., 2021). loxP511 sites are 
                     compatible with each other, but incompatible with wild-type
                     loxP sites."
     enhancer        1257..1560
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1561..1764
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     regulatory      1803..1812
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1809..1811,1812..1814,1815..2528)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1809..1811,1812..1814,1815..2525)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1854..1875)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2115..2134)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2462..2483
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(2536..2569)
                     /label=loxP511
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATAC) (Shaw et al., 2021). loxP511 sites are 
                     compatible with each other, but incompatible with wild-type
                     loxP sites."
     primer_bind     2596..2612
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    2687..3275
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2740..2760)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3158..3169)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3278..3294)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     LTR             3804..3984
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(4046..4634)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4126..4145)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4805..5596,5597..5665))
                     /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     5428..5447
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5666..5770)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5845..5864)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        6036..6415
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6417..6615
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     LTR             6633..6813
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    6860..6985
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    join(7482..7563,1..152)
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
ORIGIN
        1 caattattgt ctggtatagt gcagcagcag aacaatttgc tgagggctat tgaggcgcaa
       61 cagcatctgt tgcaactcac agtctggggc atcaagcagc tccaggcaag aatcctggct
      121 gtggaaagat acctaaagga tcaacagctc ctggggattt ggggttgctc tggaaaactc
      181 atttgcacca ctgctgtgcc ttggaatgct agttggagta ataaatctct ggaacagatt
      241 tggaatcaca cgacctggat ggagtgggac agagaaatta acaattacac aagcttaata
      301 cactccttaa ttgaagaatc gcaaaaccag caagaaaaga atgaacaaga attattggaa
      361 ttagataaat gggcaagttt gtggaattgg tttaacataa caaattggct gtggtatata
      421 aaattattca taatgatagt aggaggcttg gtaggtttaa gaatagtttt tgctgtactt
      481 tctatagtga atagagttag gcagggatat tcaccattat cgtttcagac ccacctccca
      541 accccgaggg gacccgacag gcccgaagga atagaagaag aaggtggaga gagagacaga
      601 gacagatcca ttcgattagt gaacggatcg gcactgcgtg cgccaattct gcagacaaat
      661 ggcagtattc atccacaatt ttaaaagaaa aggggggatt ggggggtaca gtgcagggga
      721 aagaatagta gacataatag caacagacat acaaactaaa gaattacaaa aacaaattac
      781 aaaaattcaa aattttcggg tttattacag ggacagcaga gatccagttt ggttagtacc
      841 gggcccgctc tagagatccg acgcgccatc tctaggcccg cgccggcccc ctcgcacaga
      901 cttgtgggag aagctcggct actcccctgc cccggttaat ttgcatataa tatttcctag
      961 taactataga ggcttaatgt gcgataaaag acagataatc tgttcttttt aatactagct
     1021 acattttaca tgataggctt ggatttctat aagagataca aatactaaat tattatttta
     1081 aaaaacagca caaaaggaaa ctcaccctaa ctgtaaagta attataactt cgtatagtat
     1141 aaattatacg aagttataag ccttggtttt ttgaattccg tattaccgcc atgcattagt
     1201 tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt
     1261 acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg
     1321 tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg
     1381 gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt
     1441 acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg
     1501 accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg
     1561 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
     1621 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
     1681 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
     1741 tgggaggtct atataagcag agctggttta gtgaaccgtc agatccgcta gcgctaccgg
     1801 tcgccaccat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg
     1861 agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg
     1921 ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct
     1981 ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc taccccgacc
     2041 acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca
     2101 ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg
     2161 acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc
     2221 tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcatg gccgacaagc
     2281 agaagaacgg catcaaggtg aacttcaaga tccgccacaa catcgaggac ggcagcgtgc
     2341 agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg
     2401 acaaccacta cctgagcacc cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc
     2461 acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt
     2521 acaagtagcg gccgcataac ttcgtatagt ataaattata cgaagttata agccttgtta
     2581 acgcgcggtg accctcgagg tcgacggtat cgataagctc gcttcacgag atcatgttta
     2641 agggttccgg ttccactagg tacaattcga tatcaagctt atcgataatc aacctctgga
     2701 ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg
     2761 tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt
     2821 ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag
     2881 gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc
     2941 caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga
     3001 actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa
     3061 ttccgtggtg ttgtcgggga aatcatcgtc ctttccttgg ctgctcgcct gtgttgccac
     3121 ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct
     3181 tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca
     3241 gacgagtcgg atctcccttt gggccgcctc cccgcatcga taccgtcgac ctcgatcgag
     3301 acctagaaaa acatggagca atcacaagta gcaatacagc agctaccaat gctgattgtg
     3361 cctggctaga agcacaagag gaggaggagg tgggttttcc agtcacacct caggtacctt
     3421 taagaccaat gacttacaag gcagctgtag atcttagcca ctttttaaaa gaaaaggggg
     3481 gactggaagg gctaattcac tcccaacgaa gacaagatat ccttgatctg tggatctacc
     3541 acacacaagg ctacttccct gattggcaga actacacacc agggccaggg atcagatatc
     3601 cactgacctt tggatggtgc tacaagctag taccagttga gcaagagaag gtagaagaag
     3661 ccaatgaagg agagaacacc cgcttgttac accctgtgag cctgcatggg atggatgacc
     3721 cggagagaga agtattagag tggaggtttg acagccgcct agcatttcat cacatggccc
     3781 gagagctgca tccggactgt actgggtctc tctggttaga ccagatctga gcctgggagc
     3841 tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct tgagtgcttc
     3901 aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc agaccctttt
     3961 agtcagtgtg gaaaatctct agcagcatgt gagcaaaagg ccagcaaaag gccaggaacc
     4021 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     4081 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     4141 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     4201 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     4261 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     4321 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     4381 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     4441 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     4501 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     4561 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     4621 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     4681 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     4741 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     4801 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     4861 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     4921 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     4981 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     5041 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     5101 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     5161 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     5221 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     5281 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     5341 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     5401 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     5461 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     5521 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     5581 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     5641 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     5701 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     5761 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt cgacggatcg ggagatctcc
     5821 cgatccccta tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagta
     5881 tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc gcgagcaaaa tttaagctac
     5941 aacaaggcaa ggcttgaccg acaattgcat gaagaatctg cttagggtta ggcgttttgc
     6001 gctgcttcgc gatgtacggg ccagatatac gcgttgacat tgattattga ctagttatta
     6061 atagtaatca attacggggt cattagttca tagcccatat atggagttcc gcgttacata
     6121 acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat
     6181 aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga
     6241 gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc
     6301 ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt
     6361 atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtgat
     6421 gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg gatttccaag
     6481 tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc
     6541 aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg tacggtggga
     6601 ggtctatata agcagcgcgt tttgcctgta ctgggtctct ctggttagac cagatctgag
     6661 cctgggagct ctctggctaa ctagggaacc cactgcttaa gcctcaataa agcttgcctt
     6721 gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag agatccctca
     6781 gaccctttta gtcagtgtgg aaaatctcta gcagtggcgc ccgaacaggg acttgaaagc
     6841 gaaagggaaa ccagaggagc tctctcgacg caggactcgg cttgctgaag cgcgcacggc
     6901 aagaggcgag gggcggcgac tggtgagtac gccaaaaatt ttgactagcg gaggctagaa
     6961 ggagagagat gggtgcgaga gcgtcagtat taagcggggg agaattagat cgcgatggga
     7021 aaaaattcgg ttaaggccag ggggaaagaa aaaatataaa ttaaaacata tagtatgggc
     7081 aagcagggag ctagaacgat tcgcagttaa tcctggcctg ttagaaacat cagaaggctg
     7141 tagacaaata ctgggacagc tacaaccatc ccttcagaca ggatcagaag aacttagatc
     7201 attatataat acagtagcaa ccctctattg tgtgcatcaa aggatagaga taaaagacac
     7261 caaggaagct ttagacaaga tagaggaaga gcaaaacaaa agtaagacca ccgcacagca
     7321 agcggccggc cgctgatctt cagacctgga ggaggagata tgagggacaa ttggagaagt
     7381 gaattatata aatataaagt agtaaaaatt gaaccattag gagtagcacc caccaaggca
     7441 aagagaagag tggtgcagag agaaaaaaga gcagtgggaa taggagcttt gttccttggg
     7501 ttcttgggag cagcaggaag cactatgggc gcagcgtcaa tgacgctgac ggtacaggcc
     7561 aga
//
