LOCUS       sequence-183380-        8868 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Retroviral vector encoding Cre-dependent expression of 
            EYFP-P2A-elfn1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8868)
  TITLE     Marin lab unpublished plasmids
REFERENCE   2  (bases 1 to 8868)
  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_elfn1
FEATURES             Location/Qualifiers
     source          1..8868
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        91..470
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        471..674
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     625..645
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             675..850
                     /label=5' LTR (truncated)
                     /note="truncated long terminal repeat from Moloney murine 
                     sarcoma virus"
     misc_feature    913..1112
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1313..1729
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1659..1681
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1697..1713
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     protein_bind    complement(2091..2124)
                     /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(2175..2208)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     regulatory      2835..2844
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(4713..4769)
                     /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(4785..5495,5496..5498,5499..5501))
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4827..4848)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     5176..5195
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5435..5456
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      5498..5507
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    5519..5552
                     /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    5603..5636
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    5646..6234
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5699..5719)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6117..6128)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             6429..6898
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(7028..7045)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7199..7787)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7279..7298)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7947..8738,8739..8807))
                     /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     8570..8589
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 tcaatattgg ctattggcca ttgcatacgt tgtatccata tcataatatg tacatttata
       61 ttggctcatg tccaacatta ccgccatgtt gacattgatt attgactagt tattaatagt
      121 aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta
      181 cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga
      241 cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt
      301 tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta
      361 ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg
      421 actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg gtgatgcggt
      481 tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc
      541 accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat
      601 gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct
      661 atataagcag agctcaataa aagagcccac aacccctcac tcggggcgcc agtcctccga
      721 ttgactgagt cgcccgggta cccgtattcc caataaagcc tcttgctgtt tgcatccgaa
      781 tcgtggtctc gctgttcctt gggagggtct cctctgagtg attgactacc cacgacgggg
      841 gtctttcatt tgggggctcg tccgggattt ggagacccct gcccagggac caccgaccca
      901 ccaccgggag gtaagctggc cagcaactta tctgtgtctg tccgattgtc tagtgtctat
      961 gtttgatgtt atgcgcctgc gtctgtacta gttagctaac tagctctgta tctggcggac
     1021 ccgtggtgga actgacgagt tctgaacacc cggccgcaac cctgggagac gtcccaggga
     1081 ctttgggggc cgtttttgtg gcccgacctg aggaagggag tcgatgtgga atccgacccc
     1141 gtcaggatat gtggttctgg taggagacga gaacctaaaa cagttcccgc ctccgtctga
     1201 atttttgctt tcggtttgga accgaagccg cgcgtcttgt ctgctgcagc gctgcagcat
     1261 cgttctgtgt tgtctctgtc tgactgtgtt tctgtatttg tctgaaaatt agggccagac
     1321 tgttaccact cccttaagtt tgaccttagg tcactggaaa gatgtcgagc ggatcgctca
     1381 caaccagtcg gtagatgtca agaagagacg ttgggttacc ttctgctctg cagaatggcc
     1441 aacctttaac gtcggatggc cgcgagacgg cacctttaac cgagacctca tcacccaggt
     1501 taagatcaag gtcttttcac ctggcccgca tggacaccca gaccaggtcc cctacatcgt
     1561 gacctgggaa gccttggctt ttgacccccc tccctgggtc aagccctttg tacaccctaa
     1621 gcctccgcct cctcttcctc catccgcccc gtctctcccc cttgaacctc ctcgttcgac
     1681 cccgcctcga tcctcccttt atccagccct cactccttct ctaggcgccg gaattcgatg
     1741 tcgactttag taccgcggac agagccttcg cccccgctgc cggcgcgcgc caccacctcc
     1801 ccagcaccaa aggcgggctg acgtcactct ccagccctcc ccaaactccc ctacctcacc
     1861 gccttggtcg cgtccgtgca gcggtgagtc cagtcgggcc gcaccacaag aggtgcaaga
     1921 taggggggtg caggcgcgac catacgctct gcggcggcag agcctcagcg ctgcctcagt
     1981 ctgcagcggg cagcagagga gtcgcgtcgt gccagagagc gccgccgtgc tcctgagccc
     2041 cttgcgctcc gcccccgcgg cccaccgacc cactgcccct tggatccgga ataacttcgt
     2101 ataggatact ttatacgaag ttatgcagaa tggtagctgg attgtagctg ctattagcaa
     2161 tatgaaacct cttaataact tcgtatagca tacattatac gaagttattt aattaatcag
     2221 gacttgtgct gggcagacac gcctttccag tagtctagga tgtcatgcag gtcctcgtct
     2281 ttggcaaact gaaccttctt gcgcagggcg tggccagccg ccatgaactc cgcatcatcc
     2341 ttgtgtctcc gtcggctcag tctgagacgc tcccagatgc ttggggatgc cctgcaggtg
     2401 tactcggggc tggatgaata gctcaggttg tggtactgtg gggacagctg cgagtagacg
     2461 aggtctcggg gtcgtggccg ggtcagaggc tccaggatgg atgccttgcg gccacccaag
     2521 ccctcgtgag tgggtggagg tggtggaggg gcaggtggct cggcagggtg agagccgggg
     2581 tatgagtgcc gatgctcgcc gtactggcga ctcttgtcgg cctccgcccg cagcacagtg
     2641 gccgcgggtg tcacagtgag gatggtctcg ggcacaggtg agcttttttc aatgtacttg
     2701 gtctcggtgg ctggggcctt atgtgtgccg gtggcctcag cccggaaggt gcgagggctg
     2761 cgtgcagaac cgctggatga agtgctggcc ctcgggggcc cgggcgcagc ctccacactg
     2821 tggtgccgct gcaggccacc atggccaaag gcatctttgt atacagggga caggaagctg
     2881 tgcttgctgg ccagaggctc ggacagcagc accagctgcg gctcagccgc agacacggcc
     2941 cctgatttgg cgccctggaa ggaggtggac tcggacttga gggcgtcgat gcagttgtta
     3001 atgatctggt tgactctgtc cacttccttg gcaatggtgg agatctccgc tactgagctc
     3061 tggggttcac ctggccggct cagctcacag cctctgcgct cctgcggctc ccccgtgcgc
     3121 acttcaatgt agttcccctt ggtggccttg ggggtctcgc tgctctccac cagcttatac
     3181 tgctccacgt cactggtggc ggctggcagg tagggtattc gagtcacagc ctcagggcca
     3241 agcagcggac cctgggtcag gggagccagg ccaggcgcct caatctcagg accatactta
     3301 agctcgatga tggttttttt caagctgcca gctgccgctg ccacagcctt cttgtgtttc
     3361 tcctcttgtc gcctccgctt gcgcaggcag taatagacag cgcccagaac cagtaccata
     3421 ccaaagaggc agcccaggat ggtcatgatg tagtgggtgg ctgtggaggg gctgggcacg
     3481 gggcccggtg ggctgggcgg cttgggcagg cagatggtaa ggcaggtgtg gttatgatgg
     3541 gtgccggagc tggtagagac cacgcagtat gtgtagttgg tgagtgtgta aagattggtg
     3601 aggcggatct cctcctgagg ctgggtcagc ttggacacgg tgaacgattt gctgttattg
     3661 tactgctcca gcgtgtacat gcggttgaac gggctcggca gctgcaccat gatggtggcc
     3721 gaattttggg tcagctgctt gaccttcatg gagggacggg cctctgtctg agggaccagc
     3781 gtagtcagga tcaccagcgg cgtggtgcca tctccggaga agcactcgtc atcagcgcag
     3841 ggcatgtcac taggctctgg tggaggcgga ggtggcgaat gtcccggctg tgaccggcct
     3901 ggcaccggac ggggtgggcc aagcacctca gcagtgtagg agccctctgt gcacacagac
     3961 tgaagcttgc taaggatgct acgttggtgg ccgtggcggc cctggcccag gaggaagtag
     4021 cctgcgtaga caggaggaga ctcacactgc acccggtcat gggtctgtgt ggcgttggtg
     4081 aaggcagcca gccagcgcag gaaacccagc agctcacagg agcagtagaa agggttgctg
     4141 tagatctcac acaccgagag cttggtcagg cctgcgaagg tgccgctgcc cagctgctgg
     4201 atgcgattca tggagagatc tatgttgacg atgttgggac actcccagaa ggcgctggcc
     4261 atcaccacct cgatgaggtt ggcctgtagg tacaggtact cgagcttgct caggccccgc
     4321 agcatgccct ccgtgaggtt gcgcagccgg ttgtagccca gctgcagcac ctggaggttg
     4381 aactggccgg agaaggcccc gtcctcgatg tagccgatct cgttcttggt gaggttgagg
     4441 tatgtgaggt ttccgaagcg gctcagggag gcgtactgca cgctgcggat acggttctcg
     4501 ttaagccgca ggtccacgat ggtgttgttg atctgttggg ggatggcctc atagggcggc
     4561 tggttctggc tgcagatggc caaccacacg aagcctttgt caccctcaat cagccagcag
     4621 tcaccttggg ccagcccacc cgcgtgcagt aacgtggcgg ctgccacaag tacccacgct
     4681 gtaccccacc cgtgcccagc cattaccggt ccaggcccgg ggttttcttc aacatcacct
     4741 gcttgcttta gcagagagaa gtttgtggcg ccggatcctc tgtacttgta cagctcgtcc
     4801 atgccgagag tgatcccggc ggcggtcacg aactccagca ggaccatgtg atcgcgcttc
     4861 tcgttggggt ctttgctcag ggcggactgg tagctcaggt agtggttgtc gggcagcagc
     4921 acggggccgt cgccgatggg ggtgttctgc tggtagtggt cggcgagctg cacgctgccg
     4981 tcctcgatgt tgtggcggat cttgaagttc accttgatgc cgttcttctg cttgtcggcc
     5041 atgatataga cgttgtggct gttgtagttg tactccagct tgtgccccag gatgttgccg
     5101 tcctccttga agtcgatgcc cttcagctcg atgcggttca ccagggtgtc gccctcgaac
     5161 ttcacctcgg cgcgggtctt gtagttgccg tcgtccttga agaagatggt gcgctcctgg
     5221 acgtagcctt cgggcatggc ggacttgaag aagtcgtgct gcttcatgtg gtcggggtag
     5281 cgggcgaagc actgcaggcc gtagccgaag gtggtcacga gggtgggcca gggcacgggc
     5341 agcttgccgg tggtgcagat gaacttcagg gtcagcttgc cgtaggtggc atcgccctcg
     5401 ccctcgccgg acacgctgaa cttgtggccg tttacgtcgc cgtccagctc gaccaggatg
     5461 ggcaccaccc cggtgaacag ctcctcgccc ttgctcacca tggtggcggc gcgccgcgat
     5521 aacttcgtat aaagtatcct atacgaagtt atttgcctta acccagaaat tatcactgtt
     5581 attctttaga atggtgcaaa gaataacttc gtataatgta tgctatacga agttatgttt
     5641 aaaccaatca acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg
     5701 ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt
     5761 cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg
     5821 agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc
     5881 ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc
     5941 tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc
     6001 ggctgttggg cactgacaat tccgtggtgt tgtcggggaa gctgacgtcc tttccatggc
     6061 tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg
     6121 ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc
     6181 gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcctggaa
     6241 ttcgagctcg agcttgttaa catcgataaa ataaaagatt ttatttagtc tccagaaaaa
     6301 ggggggaatg aaagacccca cctgtaggtt tggcaagcta gcttaagtaa cgccattttg
     6361 caaggcatgg aaaaatacat aactgagaat agagaagttc agatcaaggt caggaacaga
     6421 tggaacagct gaatatgggc caaacaggat atctgtggta agcagttcct gccccggctc
     6481 agggccaaga acagatggaa cagctgaata tgggccaaac aggatatctg tggtaagcag
     6541 ttcctgcccc ggctcagggc caagaacaga tggtccccag atgcggtcca gccctcagca
     6601 gtttctagag aaccatcaga tgtttccagg gtgccccaag gacctgaaat gaccctgtgc
     6661 cttatttgaa ctaaccaatc agttcgcttc tcgcttctgt tcgcgcgctt ctgctccccg
     6721 agctcaataa aagagcccac aacccctcac tcggggcgcc agtcctccga ttgactgagt
     6781 cgcccgggta cccgtgtatc caataaaccc tcttgcagtt gcatccgact tgtggtctcg
     6841 ctgttccttg ggagggtctc ctctgagtga ttgactaccc gtcagcgggg gtctttcaca
     6901 tgcagcatgt atcaaaatta atttggtttt ttttcttaag tatttacatt aaatggccat
     6961 agttgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     7021 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     7081 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     7141 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     7201 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     7261 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     7321 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     7381 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     7441 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     7501 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     7561 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     7621 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     7681 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     7741 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     7801 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     7861 tgagattatc aaaaaggatc ttcacctaga tccttttgcg gccgcaaatc aatctaaagt
     7921 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     7981 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     8041 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     8101 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     8161 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     8221 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     8281 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     8341 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     8401 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     8461 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     8521 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     8581 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     8641 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     8701 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     8761 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     8821 caatgatccg gccattagcc atattattca ttggttatat agcataaa
//
