LOCUS       sequence-298905-        6360 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Can be used to generate retrovirus that will mark the membrane 
            (Palmitoylated-Venus) in the presence of Cre from the rat synapsin 
            promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6360)
  AUTHORS   Lim L
  TITLE     Lynette Lim Lab Plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6360)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: RV-Syn-Dio-mVenus
FEATURES             Location/Qualifiers
     source          1..6360
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(350..383)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    complement(434..467)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(join(476..1189,1190..1192,1193..1195))
                     /codon_start=1
                     /product="yellow fluorescent protein (YFP) with fast and 
                     efficient maturation (Nagai et al., 2002)"
                     /label=Venus
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(521..542)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     870..889
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1129..1150
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     protein_bind    1270..1303
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    1354..1387
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    1397..1985
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1450..1470)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(1868..1879)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             2180..2649
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(2779..2796)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2950..3538)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3030..3049)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3698..4489,4490..4558))
                     /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     4321..4340
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     enhancer        4710..5089
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5090..5293
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5244..5264
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             5294..5469
                     /label=5' LTR (truncated)
                     /note="truncated long terminal repeat from Moloney murine 
                     sarcoma virus"
     misc_feature    5532..5731
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             5932..6348
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     6278..6300
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     6316..6332
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
ORIGIN
        1 cgactttagt accgcggaca gagccttcgc ccccgctgcc ggcgcgcgcc accacctccc
       61 cagcaccaaa ggcgggctga cgtcactctc cagccctccc caaactcccc tacctcaccg
      121 ccttggtcgc gtccgtgcag cggtgagtcc agtcgggccg caccacaaga ggtgcaagat
      181 aggggggtgc aggcgcgacc atacgctctg cggcggcaga gcctcagcgc tgcctcagtc
      241 tgcagcgggc agcagaggag tcgcgtcgtg ccagagagcg ccgccgtgct cctgagcccc
      301 ttgcgctccg cccccgcggc ccaccgaccc actgcccctt ggatccggaa taacttcgta
      361 taggatactt tatacgaagt tatgcagaat ggtagctgga ttgtagctgc tattagcaat
      421 atgaaacctc ttaataactt cgtatagcat acattatacg aagttattta attaattact
      481 tgtacagctc gtccatgccg agagtgatcc cggcggcggt cacgaactcc agcaggacca
      541 tgtgatcgcg cttctcgttg gggtctttgc tcagggcgga ctggtagctc aggtagtggt
      601 tgtcgggcag cagcacgggg ccgtcgccga tgggggtgtt ctgctggtag tggtcggcga
      661 gctgcacgcc gccgtcctcg atgttgtggc ggatcttgaa gttggccttg atgccgttct
      721 tctgcttgtc ggcggtgata tagacgttgt ggctgttgta gttgtactcc agcttgtgcc
      781 ccaggatgtt gccgtcctcc ttgaagtcga tgcccttcag ctcgatgcgg ttcaccaggg
      841 tgtcgccctc gaacttcacc tcggcgcggg tcttgtagtt gccgtcgtcc ttgaagaaga
      901 tggtgcgctc ctggacgtag ccttcgggca tggcggactt gaagaagtcg tgctgcttca
      961 tgtggtcggg gtagcgggcg aagcactgca ggccgtagcc cagggtggtc acgagggtgg
     1021 gccagggcac gggcagcttg ccggtggtgc agatcagctt cagggtcagc ttgccgtagg
     1081 tggcatcgcc ctcgccctcg ccggacacgc tgaacttgtg gccgtttacg tcgccgtcca
     1141 gctcgaccag gatgggcacc accccggtga acagctcctc gcccttgctc accatgatct
     1201 tttggtcctc atcattcttt tcaacctgtt tggttcttct catacagcac agcatggtgg
     1261 cgcgccgcga taacttcgta taaagtatcc tatacgaagt tatttgcctt aacccagaaa
     1321 ttatcactgt tattctttag aatggtgcaa agaataactt cgtataatgt atgctatacg
     1381 aagttatgtt taaaccaatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat
     1441 tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca
     1501 tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc
     1561 tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc
     1621 tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt
     1681 cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg
     1741 gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga agctgacgtc
     1801 ctttccatgg ctgctcgcct gtgttgccac ctggattctg cgcgggacgt ccttctgcta
     1861 cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg
     1921 gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc
     1981 cccgcctgga attcgagctc gagcttgtta acatcgataa aataaaagat tttatttagt
     2041 ctccagaaaa aggggggaat gaaagacccc acctgtaggt ttggcaagct agcttaagta
     2101 acgccatttt gcaaggcatg gaaaaataca taactgagaa tagagaagtt cagatcaagg
     2161 tcaggaacag atggaacagc tgaatatggg ccaaacagga tatctgtggt aagcagttcc
     2221 tgccccggct cagggccaag aacagatgga acagctgaat atgggccaaa caggatatct
     2281 gtggtaagca gttcctgccc cggctcaggg ccaagaacag atggtcccca gatgcggtcc
     2341 agccctcagc agtttctaga gaaccatcag atgtttccag ggtgccccaa ggacctgaaa
     2401 tgaccctgtg ccttatttga actaaccaat cagttcgctt ctcgcttctg ttcgcgcgct
     2461 tctgctcccc gagctcaata aaagagccca caacccctca ctcggggcgc cagtcctccg
     2521 attgactgag tcgcccgggt acccgtgtat ccaataaacc ctcttgcagt tgcatccgac
     2581 ttgtggtctc gctgttcctt gggagggtct cctctgagtg attgactacc cgtcagcggg
     2641 ggtctttcac atgcagcatg tatcaaaatt aatttggttt tttttcttaa gtatttacat
     2701 taaatggcca tagttgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     2761 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     2821 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     2881 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     2941 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     3001 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     3061 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     3121 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     3181 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     3241 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     3301 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     3361 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     3421 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     3481 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     3541 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     3601 gattttggtc atgagattat caaaaaggat cttcacctag atccttttgc ggccgcaaat
     3661 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     3721 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     3781 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     3841 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     3901 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     3961 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     4021 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     4081 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     4141 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     4201 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     4261 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     4321 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     4381 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     4441 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     4501 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     4561 tcttcctttt tcaatgatcc ggccattagc catattattc attggttata tagcataaat
     4621 caatattggc tattggccat tgcatacgtt gtatccatat cataatatgt acatttatat
     4681 tggctcatgt ccaacattac cgccatgttg acattgatta ttgactagtt attaatagta
     4741 atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac
     4801 ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac
     4861 gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt
     4921 acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat
     4981 tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga
     5041 ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg tgatgcggtt
     5101 ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca
     5161 ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg
     5221 tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta
     5281 tataagcaga gctcaataaa agagcccaca acccctcact cggggcgcca gtcctccgat
     5341 tgactgagtc gcccgggtac ccgtattccc aataaagcct cttgctgttt gcatccgaat
     5401 cgtggtctcg ctgttccttg ggagggtctc ctctgagtga ttgactaccc acgacggggg
     5461 tctttcattt gggggctcgt ccgggatttg gagacccctg cccagggacc accgacccac
     5521 caccgggagg taagctggcc agcaacttat ctgtgtctgt ccgattgtct agtgtctatg
     5581 tttgatgtta tgcgcctgcg tctgtactag ttagctaact agctctgtat ctggcggacc
     5641 cgtggtggaa ctgacgagtt ctgaacaccc ggccgcaacc ctgggagacg tcccagggac
     5701 tttgggggcc gtttttgtgg cccgacctga ggaagggagt cgatgtggaa tccgaccccg
     5761 tcaggatatg tggttctggt aggagacgag aacctaaaac agttcccgcc tccgtctgaa
     5821 tttttgcttt cggtttggaa ccgaagccgc gcgtcttgtc tgctgcagcg ctgcagcatc
     5881 gttctgtgtt gtctctgtct gactgtgttt ctgtatttgt ctgaaaatta gggccagact
     5941 gttaccactc ccttaagttt gaccttaggt cactggaaag atgtcgagcg gatcgctcac
     6001 aaccagtcgg tagatgtcaa gaagagacgt tgggttacct tctgctctgc agaatggcca
     6061 acctttaacg tcggatggcc gcgagacggc acctttaacc gagacctcat cacccaggtt
     6121 aagatcaagg tcttttcacc tggcccgcat ggacacccag accaggtccc ctacatcgtg
     6181 acctgggaag ccttggcttt tgacccccct ccctgggtca agccctttgt acaccctaag
     6241 cctccgcctc ctcttcctcc atccgccccg tctctccccc ttgaacctcc tcgttcgacc
     6301 ccgcctcgat cctcccttta tccagccctc actccttctc taggcgccgg aattcgatgt
//
