LOCUS       sequence-174271-        6518 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  retroviral vector encoding cre dependent expression of GFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6518)
  AUTHORS   Lim L, Pakan JMP, Selten MM, Marques-Smith A, Llorca A, Bae SE, 
            Rochefort NL, Marin O
  TITLE     Optimization of interneuron function by direct coupling of cell 
            migration and axonal targeting.
  JOURNAL   Nat Neurosci. 2018 Jun 18. pii: 10.1038/s41593-018-0162-9. doi: 
            10.1038/s41593-018-0162-9.
  PUBMED    29915195
REFERENCE   2  (bases 1 to 6518)
  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: "Nat 
            Neurosci. 2018 Jun 18. pii: 10.1038/s41593-018-0162-9. doi: 
            10.1038/s41593-018-0162-9."
COMMENT     Sequence Label: RV-Syn-Dio-GFP
FEATURES             Location/Qualifiers
     source          1..6518
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        266..645
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        646..849
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     800..820
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             850..1025
                     /label=5' LTR (truncated)
                     /note="truncated long terminal repeat from Moloney murine 
                     sarcoma virus"
     misc_feature    1088..1287
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1488..1904
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1834..1856
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1872..1888
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     protein_bind    complement(2266..2299)
                     /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(2350..2383)
                     /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(2400..3113,3114..3116,3117..3119))
                     /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             complement(join(2403..3113,3114..3116,3117..3119))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2445..2466)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     2794..2813
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3053..3074
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     protein_bind    3136..3169
                     /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    3220..3253
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    3263..3851
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3316..3336)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3734..3745)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(4722..4739)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4893..5481)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4973..4992)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5641..6432,6433..6501))
                     /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     6264..6283
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
       61 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
      121 cctttttcaa tgatccggcc attagccata ttattcattg gttatatagc ataaatcaat
      181 attggctatt ggccattgca tacgttgtat ccatatcata atatgtacat ttatattggc
      241 tcatgtccaa cattaccgcc atgttgacat tgattattga ctagttatta atagtaatca
      301 attacggggt cattagttca tagcccatat atggagttcc gcgttacata acttacggta
      361 aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat
      421 gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg
      481 taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc ccctattgac
      541 gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt atgggacttt
      601 cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtgat gcggttttgg
      661 cagtacatca atgggcgtgg atagcggttt gactcacggg gatttccaag tctccacccc
      721 attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc aaaatgtcgt
      781 aacaactccg ccccattgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata
      841 agcagagctc aataaaagag cccacaaccc ctcactcggg gcgccagtcc tccgattgac
      901 tgagtcgccc gggtacccgt attcccaata aagcctcttg ctgtttgcat ccgaatcgtg
      961 gtctcgctgt tccttgggag ggtctcctct gagtgattga ctacccacga cgggggtctt
     1021 tcatttgggg gctcgtccgg gatttggaga cccctgccca gggaccaccg acccaccacc
     1081 gggaggtaag ctggccagca acttatctgt gtctgtccga ttgtctagtg tctatgtttg
     1141 atgttatgcg cctgcgtctg tactagttag ctaactagct ctgtatctgg cggacccgtg
     1201 gtggaactga cgagttctga acacccggcc gcaaccctgg gagacgtccc agggactttg
     1261 ggggccgttt ttgtggcccg acctgaggaa gggagtcgat gtggaatccg accccgtcag
     1321 gatatgtggt tctggtagga gacgagaacc taaaacagtt cccgcctccg tctgaatttt
     1381 tgctttcggt ttggaaccga agccgcgcgt cttgtctgct gcagcgctgc agcatcgttc
     1441 tgtgttgtct ctgtctgact gtgtttctgt atttgtctga aaattagggc cagactgtta
     1501 ccactccctt aagtttgacc ttaggtcact ggaaagatgt cgagcggatc gctcacaacc
     1561 agtcggtaga tgtcaagaag agacgttggg ttaccttctg ctctgcagaa tggccaacct
     1621 ttaacgtcgg atggccgcga gacggcacct ttaaccgaga cctcatcacc caggttaaga
     1681 tcaaggtctt ttcacctggc ccgcatggac acccagacca ggtcccctac atcgtgacct
     1741 gggaagcctt ggcttttgac ccccctccct gggtcaagcc ctttgtacac cctaagcctc
     1801 cgcctcctct tcctccatcc gccccgtctc tcccccttga acctcctcgt tcgaccccgc
     1861 ctcgatcctc cctttatcca gccctcactc cttctctagg cgccggaatt cgatgtcgac
     1921 tttagtaccg cggacagagc cttcgccccc gctgccggcg cgcgccacca cctccccagc
     1981 accaaaggcg ggctgacgtc actctccagc cctccccaaa ctcccctacc tcaccgcctt
     2041 ggtcgcgtcc gtgcagcggt gagtccagtc gggccgcacc acaagaggtg caagataggg
     2101 gggtgcaggc gcgaccatac gctctgcggc ggcagagcct cagcgctgcc tcagtctgca
     2161 gcgggcagca gaggagtcgc gtcgtgccag agagcgccgc cgtgctcctg agccccttgc
     2221 gctccgcccc cgcggcccac cgacccactg ccccttggat ccggaataac ttcgtatagg
     2281 atactttata cgaagttatg cagaatggta gctggattgt agctgctatt agcaatatga
     2341 aacctcttaa taacttcgta tagcatacat tatacgaagt tatttaatta agcggccgct
     2401 tacttgtaca gctcgtccat gccgagagtg atcccggcgg cggtcacgaa ctccagcagg
     2461 accatgtgat cgcgcttctc gttggggtct ttgctcaggg cggactgggt gctcaggtag
     2521 tggttgtcgg gcagcagcac ggggccgtcg ccgatggggg tgttctgctg gtagtggtcg
     2581 gcgagctgca cgctgccgtc ctcgatgttg tggcggatct tgaagttcac cttgatgccg
     2641 ttcttctgct tgtcggccat gatatagacg ttgtggctgt tgtagttgta ctccagcttg
     2701 tgccccagga tgttgccgtc ctccttgaag tcgatgccct tcagctcgat gcggttcacc
     2761 agggtgtcgc cctcgaactt cacctcggcg cgggtcttgt agttgccgtc gtccttgaag
     2821 aagatggtgc gctcctggac gtagccttcg ggcatggcgg acttgaagaa gtcgtgctgc
     2881 ttcatgtggt cggggtagcg gctgaagcac tgcacgccgt aggtcagggt ggtcacgagg
     2941 gtgggccagg gcacgggcag cttgccggtg gtgcagatga acttcagggt cagcttgccg
     3001 taggtggcat cgccctcgcc ctcgccggac acgctgaact tgtggccgtt tacgtcgccg
     3061 tccagctcga ccaggatggg caccaccccg gtgaacagct cctcgccctt gctcaccatg
     3121 gggcggcgcg ccgcgataac ttcgtataaa gtatcctata cgaagttatt tgccttaacc
     3181 cagaaattat cactgttatt ctttagaatg gtgcaaagaa taacttcgta taatgtatgc
     3241 tatacgaagt tatgtttaaa ccaatcaacc tctggattac aaaatttgtg aaagattgac
     3301 tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt
     3361 gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt
     3421 gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt
     3481 gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg
     3541 gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg
     3601 ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaagct
     3661 gacgtccttt ccatggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt
     3721 ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc
     3781 tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc
     3841 cgcctccccg cctggaattc gagctcgagc ttgttaacat cgataaaata aaagatttta
     3901 tttagtctcc agaaaaaggg gggaatgaaa gaccccacct gtaggtttgg caagctagct
     3961 taagtaacgc cattttgcaa ggcatggaaa aatacataac tgagaataga gaagttcaga
     4021 tcaaggtcag gaacagatgg aacagctgaa tatgggccaa acaggatatc tgtggtaagc
     4081 agttcctgcc ccggctcagg gccaagaaca gatggaacag ctgaatatgg gccaaacagg
     4141 atatctgtgg taagcagttc ctgccccggc tcagggccaa gaacagatgg tccccagatg
     4201 cggtccagcc ctcagcagtt tctagagaac catcagatgt ttccagggtg ccccaaggac
     4261 ctgaaatgac cctgtgcctt atttgaacta accaatcagt tcgcttctcg cttctgttcg
     4321 cgcgcttctg ctccccgagc tcaataaaag agcccacaac ccctcactcg gggcgccagt
     4381 cctccgattg actgagtcgc ccgggtaccc gtgtatccaa taaaccctct tgcagttgca
     4441 tccgacttgt ggtctcgctg ttccttggga gggtctcctc tgagtgattg actacccgtc
     4501 agcgggggkc ttttgcatcc gacttgtggt ctcgctgttc cttgggaggg tctcctctga
     4561 gtgattgact acccgtcagc gggggtcttt cacatgcagc atgtatcaaa attaatttgg
     4621 ttttttttct taagtattta cattaaatgg ccatagttgc attaatgaat cggccaacgc
     4681 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     4741 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     4801 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     4861 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     4921 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     4981 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     5041 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     5101 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     5161 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     5221 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     5281 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     5341 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     5401 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     5461 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     5521 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     5581 tagatccttt tgcggccgca aatcaatcta aagtatatat gagtaaactt ggtctgacag
     5641 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     5701 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     5761 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     5821 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     5881 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     5941 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     6001 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     6061 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     6121 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     6181 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     6241 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     6301 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     6361 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     6421 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     6481 acggaaatgt tgaatactca tactcttcct ttttcaat
//
