LOCUS       sequence-366336-        7612 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Retroviral vector expression cre-dependent membrane tagged Tdtomato 
            driven by the mDlx promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7612)
  TITLE     AAV vector using mDlx promoter
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7612)
  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            "Unpublished"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: p221-RV-dlx-dio-Tdtomato
FEATURES             Location/Qualifiers
     source          1..7612
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(312..329)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(483..1071)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(563..582)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1231..2022,2023..2091))
                     /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     1854..1873
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     enhancer        2243..2622
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2623..2826
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2777..2797
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             2827..3002
                     /label=5' LTR (truncated)
                     /note="truncated long terminal repeat from Moloney murine 
                     sarcoma virus"
     misc_feature    3065..3264
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             3465..3881
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     3811..3833
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     3849..3865
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     intron          4594..4726
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     4709..4728
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     protein_bind    complement(4787..4820)
                     /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(4871..4904)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(4913..6343)
                     /codon_start=1
                     /product="tandem dimeric (pseudo-monomeric) derivative of 
                     DsRed (Shaner et al., 2004)"
                     /label=tdTomato
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     LVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH
                     HLFLGHGTGSTGSGSSGTASSEDNNMAVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPY
                     EGTQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVM
                     NFEDGGLVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGV
                     LKGEIHQALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYE
                     RSEGRHHLFLYGMDELYK"
     protein_bind    6415..6448
                     /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    6499..6532
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    6542..7130
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6595..6615)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7013..7024)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             join(7325..7612,1..182)
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
ORIGIN
        1 cccgagctca ataaaagagc ccacaacccc tcactcgggg cgccagtcct ccgattgact
       61 gagtcgcccg ggtacccgtg tatccaataa accctcttgc agttgcatcc gacttgtggt
      121 ctcgctgttc cttgggaggg tctcctctga gtgattgact acccgtcagc gggggtcttt
      181 cacatgcagc atgtatcaaa attaatttgg ttttttttct taagtattta cattaaatgg
      241 ccatagttgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
      301 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
      361 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
      421 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
      481 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
      541 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
      601 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
      661 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
      721 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
      781 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
      841 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
      901 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
      961 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     1021 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     1081 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     1141 gtcatgagat tatcaaaaag gatcttcacc tagatccttt tgcggccgca aatcaatcta
     1201 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     1261 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     1321 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     1381 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     1441 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     1501 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     1561 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     1621 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     1681 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     1741 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     1801 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     1861 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     1921 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     1981 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     2041 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     2101 ttttcaatga tccggccatt agccatatta ttcattggtt atatagcata aatcaatatt
     2161 ggctattggc cattgcatac gttgtatcca tatcataata tgtacattta tattggctca
     2221 tgtccaacat taccgccatg ttgacattga ttattgacta gttattaata gtaatcaatt
     2281 acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat
     2341 ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt
     2401 cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa
     2461 actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc
     2521 aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct
     2581 acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg gttttggcag
     2641 tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct ccaccccatt
     2701 gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa atgtcgtaac
     2761 aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt ctatataagc
     2821 agagctcaat aaaagagccc acaacccctc actcggggcg ccagtcctcc gattgactga
     2881 gtcgcccggg tacccgtatt cccaataaag cctcttgctg tttgcatccg aatcgtggtc
     2941 tcgctgttcc ttgggagggt ctcctctgag tgattgacta cccacgacgg gggtctttca
     3001 tttgggggct cgtccgggat ttggagaccc ctgcccaggg accaccgacc caccaccggg
     3061 aggtaagctg gccagcaact tatctgtgtc tgtccgattg tctagtgtct atgtttgatg
     3121 ttatgcgcct gcgtctgtac tagttagcta actagctctg tatctggcgg acccgtggtg
     3181 gaactgacga gttctgaaca cccggccgca accctgggag acgtcccagg gactttgggg
     3241 gccgtttttg tggcccgacc tgaggaaggg agtcgatgtg gaatccgacc ccgtcaggat
     3301 atgtggttct ggtaggagac gagaacctaa aacagttccc gcctccgtct gaatttttgc
     3361 tttcggtttg gaaccgaagc cgcgcgtctt gtctgctgca gcgctgcagc atcgttctgt
     3421 gttgtctctg tctgactgtg tttctgtatt tgtctgaaaa ttagggccag actgttacca
     3481 ctcccttaag tttgacctta ggtcactgga aagatgtcga gcggatcgct cacaaccagt
     3541 cggtagatgt caagaagaga cgttgggtta ccttctgctc tgcagaatgg ccaaccttta
     3601 acgtcggatg gccgcgagac ggcaccttta accgagacct catcacccag gttaagatca
     3661 aggtcttttc acctggcccg catggacacc cagaccaggt cccctacatc gtgacctggg
     3721 aagccttggc ttttgacccc cctccctggg tcaagccctt tgtacaccct aagcctccgc
     3781 ctcctcttcc tccatccgcc ccgtctctcc cccttgaacc tcctcgttcg accccgcctc
     3841 gatcctccct ttatccagcc ctcactcctt ctctaggcgc cggaattcga tgtcgacgaa
     3901 ttctatacac tcacagtggt ttggcatata tttggtgaaa ttttttaagg aaaaattagt
     3961 gttggtttcg atatatggta gctttttctc taacataatt tgaataattc agcaaagccc
     4021 tactaccagc tgtacttctg cagcctcttc cattctttcc agcattataa ttttggttaa
     4081 ttttcaattt taggtcctac gtctctgcaa tttgtgtatg aataacagaa taatttccct
     4141 cttttgtttc gcctttcctg ttcctgaatc taaataaaga tggcttttta gtattaaaag
     4201 tggaagaaaa ttacaggtaa ttatctttga cggtaaaaac gctgtaatca gcgggctaca
     4261 tgaaaaatta ctctaattat ggctgcattt aagagaatgg aaaaaaacct tcttgtggat
     4321 aaaaacctta aattgtcccc aatgtctgct tcaaattgga tggcactgca gctggaggct
     4381 ttgttcagaa ttgatcctgg ggagctacga acccaaagtt tcacagtagg aagcttgata
     4441 tcgaattcct gcagcccggg ctgggcataa aagtcagggc agagccatct attgcttaca
     4501 tttgcttcta gcctgcaggt cgaggagcgc agccttccag aagcagagcg cggcgcctta
     4561 agctgcagaa gttggtcgtg aggcactggg caggtaagta tcaaggttac aagacaggtt
     4621 taaggagacc aatagaaact gggcttgtcg agacagagaa gactcttgcg tttctgatag
     4681 gcacctattg gtcttactga catccacttt gcctttctct ccacaggtgt ccactcccag
     4741 ttcaattaca gctcttaaga attcgaaata taacatcgga tccggaataa cttcgtatag
     4801 gatactttat acgaagttat gcagaatggt agctggattg tagctgctat tagcaatatg
     4861 aaacctctta ataacttcgt atagcataca ttatacgaag ttatttaatt aattacttgt
     4921 acagctcgtc catgccgtac aggaacaggt ggtggcggcc ctcggagcgc tcgtactgtt
     4981 ccacgatggt gtagtcctcg ttgtgggagg tgatgtccag cttggtgtcc acgtagtagt
     5041 agccgggcag ttgcacgggc ttcttggcca tgtagatggt cttgaactcc accaggtagt
     5101 ggccgccgtc cttcagcttc agggcctggt ggatctcgcc cttcagcacg ccgtcgcggg
     5161 ggtacaggcg ctcggtggag gcctcccagc ccatggtctt cttctgcatt acggggccgt
     5221 cgggggggaa gttggtgccg cgcatcttca ccttgtagat cagcgtgccg tcctgcaggg
     5281 aggagtcctg ggtcacggtc accagaccgc cgtcctcgaa gttcatcacg cgctcccact
     5341 tgaagccctc ggggaaggac agcttcttgt aatcggggat gtcggcgggg tgcttcacgt
     5401 acgccttgga gccgtacatg aactgggggg acaggatgtc ccaggcgaag ggcagggggc
     5461 cgcccttggt caccttcagc ttggcggtct gggtgccctc gtaggggcgg ccctcgccct
     5521 cgccctcgat ctcgaactcg tggccgttca tggagccctc catgcgcacc ttgaagcgca
     5581 tgaactcttt gatgacggcc atgttgttgt cctcggagga ggcggtgccg gagctgccgc
     5641 tgccggtgct gccggtgcca tgccccagga acaggtggtg gcggccctcg gagcgctcgt
     5701 actgttccac gatggtgtag tcctcgttgt gggaggtgat gtccagcttg gtgtccacgt
     5761 agtagtagcc gggcagttgc acgggcttct tggccatgta gatggtcttg aactccacca
     5821 ggtagtggcc gccgtccttc agcttcaggg cctggtggat ctcgcccttc agcacgccgt
     5881 cgcgggggta caggcgctcg gtggaggcct cccagcccat ggtcttcttc tgcattacgg
     5941 ggccgtcggg ggggaagttg gtgccgcgca tcttcacctt gtagatcagc gtgccgtcct
     6001 gcagggagga gtcctgggtc acggtcacca gaccgccgtc ctcgaagttc atcacgcgct
     6061 cccacttgaa gccctcgggg aaggacagct tcttgtaatc ggggatgtcg gcggggtgct
     6121 tcacgtacgc cttggagccg tacatgaact ggggggacag gatgtcccag gcgaagggca
     6181 gggggccgcc cttggtcacc ttcagcttgg cggtctgggt gccctcgtag gggcggccct
     6241 cgccctcgcc ctcgatctcg aactcgtggc cgttcatgga gccctccatg cgcaccttga
     6301 agcgcatgaa ctctttgatg acctcctcgc ccttgctcac catgatcttt tggtcctcat
     6361 cattcttttc aacctgtttg gttcttctca tacagcacag catggcgcgc cgcgataact
     6421 tcgtataaag tatcctatac gaagttattt gccttaaccc agaaattatc actgttattc
     6481 tttagaatgg tgcaaagaat aacttcgtat aatgtatgct atacgaagtt atgtttaaac
     6541 caatcaacct ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc
     6601 tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg
     6661 tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt
     6721 gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac
     6781 tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc
     6841 tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct
     6901 gttgggcact gacaattccg tggtgttgtc ggggaagctg acgtcctttc catggctgct
     6961 cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct
     7021 caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct
     7081 tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc ctggaattcg
     7141 agctcgagct tgttaacatc gataaaataa aagattttat ttagtctcca gaaaaagggg
     7201 ggaatgaaag accccacctg taggtttggc aagctagctt aagtaacgcc attttgcaag
     7261 gcatggaaaa atacataact gagaatagag aagttcagat caaggtcagg aacagatgga
     7321 acagctgaat atgggccaaa caggatatct gtggtaagca gttcctgccc cggctcaggg
     7381 ccaagaacag atggaacagc tgaatatggg ccaaacagga tatctgtggt aagcagttcc
     7441 tgccccggct cagggccaag aacagatggt ccccagatgc ggtccagccc tcagcagttt
     7501 ctagagaacc atcagatgtt tccagggtgc cccaaggacc tgaaatgacc ctgtgcctta
     7561 tttgaactaa ccaatcagtt cgcttctcgc ttctgttcgc gcgcttctgc tc
//
