LOCUS       sequence-366334-        6399 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Retroviral vector expression cre-dependent membrane tagged 
            mScarlet-T2A driven by the rat Synapsin promoter (with XhoI and PacI
            at 3'end of T2A to clone a gene of interest).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6399)
  TITLE     AAV vector using mDlx promoter
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6399)
  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: 
            "Unpublished"
COMMENT     Sequence Label: p159-RV-syn-dio-mScarlet-T2A
FEATURES             Location/Qualifiers
     source          1..6399
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(295..312)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(466..1054)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(546..565)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1214..2005,2006..2074))
                     /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     1837..1856
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     enhancer        2226..2605
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2606..2809
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2760..2780
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             2810..2985
                     /label=5' LTR (truncated)
                     /note="truncated long terminal repeat from Moloney murine 
                     sarcoma virus"
     misc_feature    3048..3247
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             3448..3864
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     3794..3816
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     3832..3848
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     protein_bind    complement(4226..4259)
                     /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(4310..4343)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(4364..4417)
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             complement(4427..5122)
                     /codon_start=1
                     /product="bright monomeric red fluorescent protein evolved 
                     from a synthetic template (Bindels et al., 2016)"
                     /label=mScarlet-I
                     /note="matures faster than mScarlet"
                     /translation="MVSKGEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFSWDILSPQFMYGSRAFIKHPADIPDYYKQSFPEGFKWERVMNFEDGG
                     AVTVTQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDGVLKGDIK
                     MALRLKDGGRYLADFKTTYKAKKPVQMPGAYNVDRKLDITSHNEDYTVVEQYERSEGRH
                     STGGMDELYK"
     primer_bind     complement(4702..4721)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     4971..4989
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     protein_bind    5197..5230
                     /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    5281..5314
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    5324..5912
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5377..5397)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5795..5806)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             join(6095..6399,1..165)
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
ORIGIN
        1 agcccacaac ccctcactcg gggcgccagt cctccgattg actgagtcgc ccgggtaccc
       61 gtgtatccaa taaaccctct tgcagttgca tccgacttgt ggtctcgctg ttccttggga
      121 gggtctcctc tgagtgattg actacccgtc agcgggggtc tttcacatgc agcatgtatc
      181 aaaattaatt tggttttttt tcttaagtat ttacattaaa tggccatagt tgcattaatg
      241 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
      301 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
      361 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
      421 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
      481 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
      541 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
      601 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
      661 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
      721 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
      781 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
      841 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
      901 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
      961 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     1021 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     1081 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     1141 aaggatcttc acctagatcc ttttgcggcc gcaaatcaat ctaaagtata tatgagtaaa
     1201 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     1261 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     1321 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     1381 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     1441 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     1501 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     1561 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     1621 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     1681 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     1741 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     1801 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     1861 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     1921 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     1981 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     2041 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tgatccggcc
     2101 attagccata ttattcattg gttatatagc ataaatcaat attggctatt ggccattgca
     2161 tacgttgtat ccatatcata atatgtacat ttatattggc tcatgtccaa cattaccgcc
     2221 atgttgacat tgattattga ctagttatta atagtaatca attacggggt cattagttca
     2281 tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc
     2341 gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat
     2401 agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt
     2461 acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc
     2521 cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta
     2581 cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg
     2641 atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt
     2701 gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac
     2761 gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctc aataaaagag
     2821 cccacaaccc ctcactcggg gcgccagtcc tccgattgac tgagtcgccc gggtacccgt
     2881 attcccaata aagcctcttg ctgtttgcat ccgaatcgtg gtctcgctgt tccttgggag
     2941 ggtctcctct gagtgattga ctacccacga cgggggtctt tcatttgggg gctcgtccgg
     3001 gatttggaga cccctgccca gggaccaccg acccaccacc gggaggtaag ctggccagca
     3061 acttatctgt gtctgtccga ttgtctagtg tctatgtttg atgttatgcg cctgcgtctg
     3121 tactagttag ctaactagct ctgtatctgg cggacccgtg gtggaactga cgagttctga
     3181 acacccggcc gcaaccctgg gagacgtccc agggactttg ggggccgttt ttgtggcccg
     3241 acctgaggaa gggagtcgat gtggaatccg accccgtcag gatatgtggt tctggtagga
     3301 gacgagaacc taaaacagtt cccgcctccg tctgaatttt tgctttcggt ttggaaccga
     3361 agccgcgcgt cttgtctgct gcagcgctgc agcatcgttc tgtgttgtct ctgtctgact
     3421 gtgtttctgt atttgtctga aaattagggc cagactgtta ccactccctt aagtttgacc
     3481 ttaggtcact ggaaagatgt cgagcggatc gctcacaacc agtcggtaga tgtcaagaag
     3541 agacgttggg ttaccttctg ctctgcagaa tggccaacct ttaacgtcgg atggccgcga
     3601 gacggcacct ttaaccgaga cctcatcacc caggttaaga tcaaggtctt ttcacctggc
     3661 ccgcatggac acccagacca ggtcccctac atcgtgacct gggaagcctt ggcttttgac
     3721 ccccctccct gggtcaagcc ctttgtacac cctaagcctc cgcctcctct tcctccatcc
     3781 gccccgtctc tcccccttga acctcctcgt tcgaccccgc ctcgatcctc cctttatcca
     3841 gccctcactc cttctctagg cgccggaatt cgatgtcgac tttagtaccg cggacagagc
     3901 cttcgccccc gctgccggcg cgcgccacca cctccccagc accaaaggcg ggctgacgtc
     3961 actctccagc cctccccaaa ctcccctacc tcaccgcctt ggtcgcgtcc gtgcagcggt
     4021 gagtccagtc gggccgcacc acaagaggtg caagataggg gggtgcaggc gcgaccatac
     4081 gctctgcggc ggcagagcct cagcgctgcc tcagtctgca gcgggcagca gaggagtcgc
     4141 gtcgtgccag agagcgccgc cgtgctcctg agccccttgc gctccgcccc cgcggcccac
     4201 cgacccactg ccccttggat ccggaataac ttcgtatagg atactttata cgaagttatg
     4261 cagaatggta gctggattgt agctgctatt agcaatatga aacctcttaa taacttcgta
     4321 tagcatacat tatacgaagt tatttaatta aatgatgctc gagagggccg ggattctctt
     4381 ccacgtctcc gcatgtcaga agacttcctc tgccctcgcc acttcccttg tacagctcgt
     4441 ccatgccgcc ggtggagtgg cggccctcgg agcgttcgta ctgttccacc acggtgtagt
     4501 cctcgttgtg ggaggtgatg tccaacttgc ggtcgacgtt gtaggcgccg ggcatctgca
     4561 cgggcttctt ggccttgtag gtggtcttga agtccgccag gtagcggccg ccgtccttca
     4621 ggcgcagggc catcttaatg tcgcccttca gcacgccgtc ctcggggtac aaccgctcgg
     4681 tggacgcttc ccagcccatt gtcttcttct gcattacggg gccgtcagga gggaagttgg
     4741 tgccgcggag cttcaccttg tagatcaggg tgccgtcctc cagggaggtg tcctgggtca
     4801 cggtcacggc gccgccgtcc tcgaagttca tcacgcgctc ccacttgaag ccctcgggga
     4861 aggactgctt atagtagtcg gggatgtcgg cggggtgctt gatgaaggcc ctggagccgt
     4921 acatgaactg aggggacagg atgtcccagg agaagggcag ggggccaccc ttggtcacct
     4981 tcagcttggc ggtctgggtg ccctcgtagg ggcggccctc gccctcgccc tcgatctcga
     5041 actcgtggcc gttcatggag ccctccatgt gcaccttgaa ccgcatgaac tccttgatca
     5101 ctgcctcgcc cttgctcacc atgatctttt ggtcctcatc attcttttca acctgtttgg
     5161 ttcttctcat acagcacagc atggtggcgc gccgcgataa cttcgtataa agtatcctat
     5221 acgaagttat ttgccttaac ccagaaatta tcactgttat tctttagaat ggtgcaaaga
     5281 ataacttcgt ataatgtatg ctatacgaag ttatgtttaa accaatcaac ctctggatta
     5341 caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg
     5401 atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc
     5461 ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca
     5521 acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac
     5581 cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact
     5641 catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc
     5701 cgtggtgttg tcggggaagc tgacgtcctt tccatggctg ctcgcctgtg ttgccacctg
     5761 gattctgcgc gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc
     5821 ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac
     5881 gagtcggatc tccctttggg ccgcctcccc gcctggaatt cgagctcgac gataaaataa
     5941 aagattttat ttagtctcca gaaaaagggg ggaatgaaag accccacctg taggtttggc
     6001 aagctagctt aagtaacgcc attttgcaag gcatggaaaa atacataact gagaatagag
     6061 aagttcagat caaggtcagg aacagatgga acagctgaat atgggccaaa caggatatct
     6121 gtggtaagca gttcctgccc cggctcaggg ccaagaacag atggaacagc tgaatatggg
     6181 ccaaacagga tatctgtggt aagcagttcc tgccccggct cagggccaag aacagatggt
     6241 ccccagatgc ggtccagccc tcagcagttt ctagagaacc atcagatgtt tccagggtgc
     6301 cccaaggacc tgaaatgacc ctgtgcctta tttgaactaa ccaatcagtt cgcttctcgc
     6361 ttctgttcgc gcgcttctgc tccccgagct caataaaag
//
