LOCUS       sequence-179955-        8832 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Retroviral vector encoding Cre-dependent expression of 
            EYFP-P2A-Mafb.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8832)
  AUTHORS   Mi D, Li Z, Lim L, Li M, Moissidis M, Yang Y, Gao T, Hu TX, Pratt T,
            Price DJ, Sestan N, Marin O
  TITLE     Early emergence of cortical interneuron diversity in the mouse 
            embryo.
  JOURNAL   Science. 2018 Feb 22. pii: science.aar6821. doi: 
            10.1126/science.aar6821.
  PUBMED    29472441
REFERENCE   2  (bases 1 to 8832)
  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: "Science. 
            2018 Feb 22. pii: science.aar6821. doi: 10.1126/science.aar6821."
COMMENT     Sequence Label: RV-Cag-Dio-EYFP-P2A-Mafb
FEATURES             Location/Qualifiers
     source          1..8832
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        135..514
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        515..718
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     LTR             719..894
                     /label=5' LTR (truncated)
                     /note="truncated long terminal repeat from Moloney murine 
                     sarcoma virus"
     misc_feature    957..1156
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1357..1773
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1703..1725
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1741..1757
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     enhancer        1787..2166
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2169..2444
                     /label=chicken beta-actin promoter
     intron          2445..3453
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     3377..3399
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     protein_bind    complement(3523..3556)
                     /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(3607..3640)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(4120..4137)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(4207..4230)
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     CDS             complement(4630..4686)
                     /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(4702..5412,5413..5415,5416..5418))
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4744..4765)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     5093..5112
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5352..5373
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      5415..5424
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    5436..5469
                     /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    5520..5553
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    5563..6151
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5616..5636)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6034..6045)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             6224..6817
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(7018..7035)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7189..7777)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7269..7288)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7955..8746,8747..8815))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     8578..8597
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 gatccggcca ttagccatat tattcattgg ttatatagca taaatcaata ttggctattg
       61 gccattgcat acgttgtatc catatcataa tatgtacatt tatattggct catgtccaac
      121 attaccgcca tgttgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc
      181 attagttcat agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc
      241 tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt
      301 aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca
      361 cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg
      421 taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca
      481 gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa
      541 tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa
      601 tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc
      661 cccattgacg caaatgggcg gtaggcatgt acggtgggag gtctatataa gcagagctca
      721 ataaaagagc ccacaacccc tcactcgggg cgccagtcct ccgattgact gagtcgcccg
      781 ggtacccgta ttcccaataa agcctcttgc tgtttgcatc cgaatcgtgg tctcgctgtt
      841 ccttgggagg gtctcctctg agtgattgac tacccacgac gggggtcttt catttggggg
      901 ctcgtccggg atttggagac ccctgcccag ggaccaccga cccaccaccg ggaggtaagc
      961 tggccagcaa cttatctgtg tctgtccgat tgtctagtgt ctatgtttga tgttatgcgc
     1021 ctgcgtctgt actagttagc taactagctc tgtatctggc ggacccgtgg tggaactgac
     1081 gagttctgaa cacccggccg caaccctggg agacgtccca gggactttgg gggccgtttt
     1141 tgtggcccga cctgaggaag ggagtcgatg tggaatccga ccccgtcagg atatgtggtt
     1201 ctggtaggag acgagaacct aaaacagttc ccgcctccgt ctgaattttt gctttcggtt
     1261 tggaaccgaa gccgcgcgtc ttgtctgctg cagcgctgca gcatcgttct gtgttgtctc
     1321 tgtctgactg tgtttctgta tttgtctgaa aattagggcc agactgttac cactccctta
     1381 agtttgacct taggtcactg gaaagatgtc gagcggatcg ctcacaacca gtcggtagat
     1441 gtcaagaaga gacgttgggt taccttctgc tctgcagaat ggccaacctt taacgtcgga
     1501 tggccgcgag acggcacctt taaccgagac ctcatcaccc aggttaagat caaggtcttt
     1561 tcacctggcc cgcatggaca cccagaccag gtcccctaca tcgtgacctg ggaagccttg
     1621 gcttttgacc cccctccctg ggtcaagccc tttgtacacc ctaagcctcc gcctcctctt
     1681 cctccatccg ccccgtctct cccccttgaa cctcctcgtt cgaccccgcc tcgatcctcc
     1741 ctttatccag ccctcactcc ttctctaggc gccggaattc gatgtcgaca ttgattattg
     1801 actagttatt aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc
     1861 cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca
     1921 ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt
     1981 caatgggtgg actatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg
     2041 ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag
     2101 tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt
     2161 accatgggtc gaggtgagcc ccacgttctg cttcactctc cccatctccc ccccctcccc
     2221 acccccaatt ttgtatttat ttatttttta attattttgt gcagcgatgg gggcgggggg
     2281 ggggggggcg cgcgccaggc ggggcggggc ggggcgaggg gcggggcggg gcgaggcgga
     2341 gaggtgcggc ggcagccaat cagagcggcg cgctccgaaa gtttcctttt atggcgaggc
     2401 ggcggcggcg gcggccctat aaaaagcgaa gcgcgcggcg ggcgggagtc gctgcgttgc
     2461 cttcgccccg tgccccgctc cgcgccgcct cgcgccgccc gccccggctc tgactgaccg
     2521 cgttactccc acaggtgagc gggcgggacg gcccttctcc tccgggctgt aattagcgct
     2581 tggtttaatg acggctcgtt tcttttctgt ggctgcgtga aagccttaaa gggctccggg
     2641 agggcccttt gtgcgggggg gagcggctcg gggggtgcgt gcgtgtgtgt gtgcgtgggg
     2701 agcgccgcgt gcggcccgcg ctgcccggcg gctgtgagcg ctgcgggcgc ggcgcggggc
     2761 tttgtgcgct ccgcgtgtgc gcgaggggag cgcggccggg ggcggtgccc cgcggtgcgg
     2821 gggggctgcg aggggaacaa aggctgcgtg cggggtgtgt gcgtgggggg gtgagcaggg
     2881 ggtgtgggcg cggcggtcgg gctgtaaccc ccccctgcac ccccctcccc gagttgctga
     2941 gcacggcccg gcttcgggtg cggggctccg tgcggggcgt ggcgcggggc tcgccgtgcc
     3001 gggcgggggg tggcggcagg tgggggtgcc gggcggggcg gggccgcctc gggccgggga
     3061 gggctcgggg gaggggcgcg gcggccccgg agcgccggcg gctgtcgagg cgcggcgagc
     3121 cgcagccatt gccttttatg gtaatcgtgc gagagggcgc agggacttcc tttgtcccaa
     3181 atctggcgga gccgaaatct gggaggcgcc gccgcacccc ctctagcggg cgcgggcgaa
     3241 gcggtgcggc gccggcagga aggaaatggg cggggagggc cttcgtgcgt cgccgcgccg
     3301 ccgtcccctt ctccatctcc agcctcgggg ctgccgcagg gggacggctg ccttcggggg
     3361 ggacggggca gggcggggtt cggcttctgg cgtgtgaccg gcggctctag agcctctgct
     3421 aaccatgttc atgccttctt ctttttccta cagctcctgg gcaacgtgct ggttgttgtg
     3481 ctgtctcatc attttggcaa agaattcgct agcggatccg gaataacttc gtataggata
     3541 ctttatacga agttatgcag aatggtagct ggattgtagc tgctattagc aatatgaaac
     3601 ctcttaataa cttcgtatag catacattat acgaagttat ttaattaatc acagaaagaa
     3661 ctcaggagag gaggggctgt cgctggtgga gcccgcctcc ctgaagccgg agttggcgag
     3721 tttctcgcac ttgaccttgt aggcgtctct ctcgcgggcc agccgggaca cctcctgctt
     3781 aagctgctcc acctgctgaa tgagctgcgt cttctcgttc tccaggtgat gtttctgctg
     3841 gacgcgttta tacctgcacg actgggcgta gccccggttc ttcagggtcc gccgcttctg
     3901 cttcaggcgg atcacctcgt ccttggtgaa gccccgcagg tggcggttca gctcacgcac
     3961 cgacatggac accagctggt catcagagaa gcggtcctcc acactaccgt tgccgcccgc
     4021 agccgtcgcc gcggctgttg cgtgcggtcc cggccccggg tggctagtgg gtagctgttg
     4081 cgcgggactg gcagcgctgg acggcggggg cgacgcttgg tgatgatggt gatggtgcgg
     4141 gtgagcgtgc tggcccaggt catcgtgagt cacacctgct cctgggtacc cgtggtgcgg
     4201 atgagggtgg tggtggtggt gatggtggtg cgcactacgg aagccgtcga agctctgcag
     4261 cggctggggc actgggtgcg aaccgatgag cgcctccacc gcgtcctcgg gcgtcaggtt
     4321 gagtgcctcg gggttcatct gctggtagtt gctcgccatc cagtacaggt cctcgagatg
     4381 ggtcttcggt tcagtcggac tgaagctggg agaagaaggc acggagctgc acggagtgct
     4441 gagcggggtg gacgacaccg agccagcagg ctgcaggcgt gtgcatggcc ggcccggacg
     4501 ctccgcgcgc cccaggggct ccttcttcac gtcgaacttg agaaggtcga agtcgttgac
     4561 gtactccatg gccagtgggc tggtgggcag ctcttgcccc atgctcagct ccgcggccat
     4621 taccggtcca ggcccggggt tttcttcaac atcacctgct tgctttagca gagagaagtt
     4681 tgtggcgccg gatcctctgt acttgtacag ctcgtccatg ccgagagtga tcccggcggc
     4741 ggtcacgaac tccagcagga ccatgtgatc gcgcttctcg ttggggtctt tgctcagggc
     4801 ggactggtag ctcaggtagt ggttgtcggg cagcagcacg gggccgtcgc cgatgggggt
     4861 gttctgctgg tagtggtcgg cgagctgcac gctgccgtcc tcgatgttgt ggcggatctt
     4921 gaagttcacc ttgatgccgt tcttctgctt gtcggccatg atatagacgt tgtggctgtt
     4981 gtagttgtac tccagcttgt gccccaggat gttgccgtcc tccttgaagt cgatgccctt
     5041 cagctcgatg cggttcacca gggtgtcgcc ctcgaacttc acctcggcgc gggtcttgta
     5101 gttgccgtcg tccttgaaga agatggtgcg ctcctggacg tagccttcgg gcatggcgga
     5161 cttgaagaag tcgtgctgct tcatgtggtc ggggtagcgg gcgaagcact gcaggccgta
     5221 gccgaaggtg gtcacgaggg tgggccaggg cacgggcagc ttgccggtgg tgcagatgaa
     5281 cttcagggtc agcttgccgt aggtggcatc gccctcgccc tcgccggaca cgctgaactt
     5341 gtggccgttt acgtcgccgt ccagctcgac caggatgggc accaccccgg tgaacagctc
     5401 ctcgcccttg ctcaccatgg tggcggcgcg ccgcgataac ttcgtataaa gtatcctata
     5461 cgaagttatt tgccttaacc cagaaattat cactgttatt ctttagaatg gtgcaaagaa
     5521 taacttcgta taatgtatgc tatacgaagt tatgtttaaa ccaatcaacc tctggattac
     5581 aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga
     5641 tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc
     5701 tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa
     5761 cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc
     5821 acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc
     5881 atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc
     5941 gtggtgttgt cggggaagct gacgtccttt ccatggctgc tcgcctgtgt tgccacctgg
     6001 attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct
     6061 tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg
     6121 agtcggatct ccctttgggc cgcctccccg cctggaattc gagctcgagc ttgttaacat
     6181 cgataaaata aaagatttta tttagtctcc agaaaaaggg gggaatgaaa gaccccacct
     6241 gtaggtttgg caagctagct taagtaacgc cattttgcaa ggcatggaaa aatacataac
     6301 tgagaataga gaagttcaga tcaaggtcag gaacagatgg aacagctgaa tatgggccaa
     6361 acaggatatc tgtggtaagc agttcctgcc ccggctcagg gccaagaaca gatggaacag
     6421 ctgaatatgg gccaaacagg atatctgtgg taagcagttc ctgccccggc tcagggccaa
     6481 gaacagatgg tccccagatg cggtccagcc ctcagcagtt tctagagaac catcagatgt
     6541 ttccagggtg ccccaaggac ctgaaatgac cctgtgcctt atttgaacta accaatcagt
     6601 tcgcttctcg cttctgttcg cgcgcttctg ctccccgagc tcaataaaag agcccacaac
     6661 ccctcactcg gggcgccagt cctccgattg actgagtcgc ccgggtaccc gtgtatccaa
     6721 taaaccctct tgcagttgca tccgacttgt ggtctcgctg ttccttggga gggtctcctc
     6781 tgagtgattg actacccgtc agcgggggtc tttcatttcc gacttgtggt ctcgctgcct
     6841 tgggagggtc tcctctgagt gattgactac ccgtcagcgg gggtcttcac atgcagcatg
     6901 tatcaaaatt aatttggttt tttttcttaa gtatttacat taaatggcca tagttgcatt
     6961 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tggcgctctt ccgcttcctc
     7021 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     7081 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     7141 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     7201 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     7261 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     7321 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     7381 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     7441 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     7501 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     7561 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     7621 cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     7681 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     7741 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     7801 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     7861 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agtttgcggc cgcaaatcaa
     7921 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     7981 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     8041 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     8101 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     8161 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     8221 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     8281 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     8341 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     8401 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     8461 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     8521 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca acacgggata
     8581 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     8641 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     8701 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     8761 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     8821 tcctttttca at
//
