LOCUS       sequence-193481-        7983 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Retroviral vector encoding Cre-dependent expression of 
            EYFP-P2A-Neto1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7983)
  TITLE     Marin lab unpublished plasmids
REFERENCE   2  (bases 1 to 7983)
  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"
COMMENT     Sequence Label: RV-Syn-Dio-yfp_2a_neto1
FEATURES             Location/Qualifiers
     source          1..7983
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(617..1408,1409..1477))
                     /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     1240..1259
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     enhancer        1629..2008
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2009..2212
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2163..2183
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             2213..2388
                     /label=5' LTR (truncated)
                     /note="truncated long terminal repeat from Moloney murine 
                     sarcoma virus"
     misc_feature    2451..2650
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             2851..3267
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     3197..3219
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     3235..3251
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     protein_bind    complement(3629..3662)
                     /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(3713..3746)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(5366..5422)
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             complement(join(5438..6148,6149..6151,6152..6154))
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(5480..5501)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     5829..5848
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     6088..6109
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      6151..6160
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    6172..6205
                     /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    6256..6289
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    6299..6887
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6352..6372)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6770..6781)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             7082..7551
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(7681..7698)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(7852..7983,1..457))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7932..7951)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
       61 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
      121 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
      181 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
      241 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
      301 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
      361 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
      421 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
      481 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
      541 aaaaaggatc ttcacctaga tccttttgcg gccgcaaatc aatctaaagt atatatgagt
      601 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
      661 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
      721 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
      781 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
      841 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
      901 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
      961 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     1021 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     1081 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     1141 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     1201 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     1261 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     1321 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     1381 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     1441 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatgatccg
     1501 gccattagcc atattattca ttggttatat agcataaatc aatattggct attggccatt
     1561 gcatacgttg tatccatatc ataatatgta catttatatt ggctcatgtc caacattacc
     1621 gccatgttga cattgattat tgactagtta ttaatagtaa tcaattacgg ggtcattagt
     1681 tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg
     1741 accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc
     1801 aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc
     1861 agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg
     1921 gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat
     1981 ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca tcaatgggcg
     2041 tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag
     2101 tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact ccgccccatt
     2161 gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag ctcaataaaa
     2221 gagcccacaa cccctcactc ggggcgccag tcctccgatt gactgagtcg cccgggtacc
     2281 cgtattccca ataaagcctc ttgctgtttg catccgaatc gtggtctcgc tgttccttgg
     2341 gagggtctcc tctgagtgat tgactaccca cgacgggggt ctttcatttg ggggctcgtc
     2401 cgggatttgg agacccctgc ccagggacca ccgacccacc accgggaggt aagctggcca
     2461 gcaacttatc tgtgtctgtc cgattgtcta gtgtctatgt ttgatgttat gcgcctgcgt
     2521 ctgtactagt tagctaacta gctctgtatc tggcggaccc gtggtggaac tgacgagttc
     2581 tgaacacccg gccgcaaccc tgggagacgt cccagggact ttgggggccg tttttgtggc
     2641 ccgacctgag gaagggagtc gatgtggaat ccgaccccgt caggatatgt ggttctggta
     2701 ggagacgaga acctaaaaca gttcccgcct ccgtctgaat ttttgctttc ggtttggaac
     2761 cgaagccgcg cgtcttgtct gctgcagcgc tgcagcatcg ttctgtgttg tctctgtctg
     2821 actgtgtttc tgtatttgtc tgaaaattag ggccagactg ttaccactcc cttaagtttg
     2881 accttaggtc actggaaaga tgtcgagcgg atcgctcaca accagtcggt agatgtcaag
     2941 aagagacgtt gggttacctt ctgctctgca gaatggccaa cctttaacgt cggatggccg
     3001 cgagacggca cctttaaccg agacctcatc acccaggtta agatcaaggt cttttcacct
     3061 ggcccgcatg gacacccaga ccaggtcccc tacatcgtga cctgggaagc cttggctttt
     3121 gacccccctc cctgggtcaa gccctttgta caccctaagc ctccgcctcc tcttcctcca
     3181 tccgccccgt ctctccccct tgaacctcct cgttcgaccc cgcctcgatc ctccctttat
     3241 ccagccctca ctccttctct aggcgccgga attcgatgtc gactttagta ccgcggacag
     3301 agccttcgcc cccgctgccg gcgcgcgcca ccacctcccc agcaccaaag gcgggctgac
     3361 gtcactctcc agccctcccc aaactcccct acctcaccgc cttggtcgcg tccgtgcagc
     3421 ggtgagtcca gtcgggccgc accacaagag gtgcaagata ggggggtgca ggcgcgacca
     3481 tacgctctgc ggcggcagag cctcagcgct gcctcagtct gcagcgggca gcagaggagt
     3541 cgcgtcgtgc cagagagcgc cgccgtgctc ctgagcccct tgcgctccgc ccccgcggcc
     3601 caccgaccca ctgccccttg gatccggaat aacttcgtat aggatacttt atacgaagtt
     3661 atgcagaatg gtagctggat tgtagctgct attagcaata tgaaacctct taataacttc
     3721 gtatagcata cattatacga agttatttaa ttaattagac cctagttgtg ttgtattcag
     3781 attcatgttt gcttagagac ccaatgaggc agaaccgctg gacagatttt tcgtgtctgg
     3841 atagcctgtg gcttgtggtg ggcacctcct caatctcgtc gatgtcacag gcatcggcgg
     3901 catcttgtga gtagttgtgt ttcatgacca ggatgttcct tctgttcacg ggtgggatga
     3961 ggggcttgac tggctgtgtg ggtatctctg ccaaaatgga agcgtccctt gtgctgaggt
     4021 tactgcgact gcctttcgcg ctggacagtt gggatccaca gtgatggtca tggatgcatt
     4081 tggaagatga tctcctcagt ttatggtaat tttcaaagtc ttccgccacg tctgcaaagt
     4141 cggctgttgc tccagttcct ctgagcgtac ataactcata gtggggaggc tcaaacacct
     4201 cctggaaaac ggtctggtca aagtctgact tcctttggac gtatttttta cgaggctgtt
     4261 tgatctgtac aatgacagaa acgataataa ggatgatcac aatgcaggaa gtcacgccaa
     4321 tgaccgtccc actggtgttg gtcagctgat ccagcaggct ggttttcctc ttttctttac
     4381 agtgattttc atcccaggga tatacacagt tctggagccc attgcagact agcgtgttgt
     4441 taatacacat gttactgtgg cagaagaatg tgttgccttc acagggaggt tcttgaaagg
     4501 atgtgaagag catctgaaac ctgctgttac ggctaccctc gtctgcccac atgcggatca
     4561 ccccaaggcc tgtgcgcagc atgacgtcat tagccactgt gctgcagaac ttggccttca
     4621 aatcctccac ggagctgctt ccgtcataga ccgccacaaa gtttcttttg cactcatttg
     4681 aattctgcat ctcatagtcc aagaaacgta agtaaatctt ggatcgtggt ggggctcgga
     4741 tgtaccattt gcaatccacg gcttcactgg cagaagcttt gccttccttc agaatctgta
     4801 tggactccac aattccttcc gggccgccca tctcaaactc acacgctggc aatggtttca
     4861 aaactccaag gtccttaaag tcgggatcag gtgtgaaatt atatcgagct gaaaatccca
     4921 tagactccag ctcgccatca gcaaaaaatt taatccacag aaatcttcca ctagatttta
     4981 ttacaggtgg attctgttgt ccacagaacc ttccaattat tggagaaaag ccaaagggtc
     5041 catctcgaac ttcaatatga tcaaatttgc actcccaaga tggttcgatt gagtattttt
     5101 catcaaagta aagttcaatg cactgccttg gggcagcttc tatgatgtag acacactctc
     5161 ggtctggggg atatttgctg ggataattgg gagacgtaaa gacacctccc tctgcatgct
     5221 ttgtccacgt tccacactgc actgacttct gtgtttctgg ggtgatttgt ttttctgttc
     5281 ctttcttggt tgctccagaa gaatggagga tgattaaact tgctataatg tgaaacaaac
     5341 tgcgtccata gatcataacc ggtccaggcc cggggttttc ttcaacatca cctgcttgct
     5401 ttagcagaga gaagtttgtg gcgccggatc ctctgtactt gtacagctcg tccatgccga
     5461 gagtgatccc ggcggcggtc acgaactcca gcaggaccat gtgatcgcgc ttctcgttgg
     5521 ggtctttgct cagggcggac tggtagctca ggtagtggtt gtcgggcagc agcacggggc
     5581 cgtcgccgat gggggtgttc tgctggtagt ggtcggcgag ctgcacgctg ccgtcctcga
     5641 tgttgtggcg gatcttgaag ttcaccttga tgccgttctt ctgcttgtcg gccatgatat
     5701 agacgttgtg gctgttgtag ttgtactcca gcttgtgccc caggatgttg ccgtcctcct
     5761 tgaagtcgat gcccttcagc tcgatgcggt tcaccagggt gtcgccctcg aacttcacct
     5821 cggcgcgggt cttgtagttg ccgtcgtcct tgaagaagat ggtgcgctcc tggacgtagc
     5881 cttcgggcat ggcggacttg aagaagtcgt gctgcttcat gtggtcgggg tagcgggcga
     5941 agcactgcag gccgtagccg aaggtggtca cgagggtggg ccagggcacg ggcagcttgc
     6001 cggtggtgca gatgaacttc agggtcagct tgccgtaggt ggcatcgccc tcgccctcgc
     6061 cggacacgct gaacttgtgg ccgtttacgt cgccgtccag ctcgaccagg atgggcacca
     6121 ccccggtgaa cagctcctcg cccttgctca ccatggtggc ggcgcgccgc gataacttcg
     6181 tataaagtat cctatacgaa gttatttgcc ttaacccaga aattatcact gttattcttt
     6241 agaatggtgc aaagaataac ttcgtataat gtatgctata cgaagttatg tttaaaccaa
     6301 tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc
     6361 ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat
     6421 ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg
     6481 gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg
     6541 ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat
     6601 tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt
     6661 gggcactgac aattccgtgg tgttgtcggg gaagctgacg tcctttccat ggctgctcgc
     6721 ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa
     6781 tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg
     6841 ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcctg gaattcgagc
     6901 tcgagcttgt taacatcgat aaaataaaag attttattta gtctccagaa aaagggggga
     6961 atgaaagacc ccacctgtag gtttggcaag ctagcttaag taacgccatt ttgcaaggca
     7021 tggaaaaata cataactgag aatagagaag ttcagatcaa ggtcaggaac agatggaaca
     7081 gctgaatatg ggccaaacag gatatctgtg gtaagcagtt cctgccccgg ctcagggcca
     7141 agaacagatg gaacagctga atatgggcca aacaggatat ctgtggtaag cagttcctgc
     7201 cccggctcag ggccaagaac agatggtccc cagatgcggt ccagccctca gcagtttcta
     7261 gagaaccatc agatgtttcc agggtgcccc aaggacctga aatgaccctg tgccttattt
     7321 gaactaacca atcagttcgc ttctcgcttc tgttcgcgcg cttctgctcc ccgagctcaa
     7381 taaaagagcc cacaacccct cactcggggc gccagtcctc cgattgactg agtcgcccgg
     7441 gtacccgtgt atccaataaa ccctcttgca gttgcatccg acttgtggtc tcgctgttcc
     7501 ttgggagggt ctcctctgag tgattgacta cccgtcagcg ggggtctttc acatgcagca
     7561 tgtatcaaaa ttaatttggt tttttttctt aagtatttac attaaatggc catagttgca
     7621 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     7681 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     7741 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     7801 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     7861 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     7921 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     7981 tcc
//
