LOCUS       sequence-458839-        7976 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Retrovirus driving overexpression of VEGF120 plus mCherry reporter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7976)
  AUTHORS   Miller LN, Walters AE, Denninger JK, Hanson MA, Marshall AH, 
            Johantges AC, Hosawi M, Sebring G, Rieskamp JD, Ding T, Rindani R, 
            Chen KS, Goldberg ME, Senthilvelan S, Volk A, Zhao F, Askwith C, 
            Wester JC, Kirby ED
  TITLE     Neural stem and progenitor cells support and protect adult 
            hippocampal function via vascular endothelial growth factor 
            secretion.
  JOURNAL   Mol Psychiatry. 2024 Nov 11. doi: 10.1038/s41380-024-02827-8.
  PUBMED    39528687
REFERENCE   2  (bases 1 to 7976)
  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: "Mol 
            Psychiatry. 2024 Nov 11. doi: 10.1038/s41380-024-02827-8."
COMMENT     Sequence Label: CAG-VEGF120-T2A-mCherry
FEATURES             Location/Qualifiers
     source          1..7976
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        17..396
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        397..600
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     551..571
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             601..776
                     /label=5' LTR (truncated)
                     /note="truncated long terminal repeat from Moloney murine 
                     sarcoma virus"
     misc_feature    839..1038
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1239..1655
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1585..1607
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1623..1639
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     enhancer        1669..2048
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2050..2328
                     /label=chicken beta-actin promoter
     intron          2329..3345
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     3269..3291
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     3353..3372
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             3876..3929
                     /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             3930..4637
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(4075..4093)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     4343..4362
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             4638..4667
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     misc_feature    4680..5268
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4733..4753)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5151..5162)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             5463..5932
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(6062..6079)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6233..6821)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6313..6332)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6981..7772,7773..7841))
                     /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     7604..7623
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 acattaccgc catgttgaca ttgattattg actagttatt aatagtaatc aattacgggg
       61 tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg
      121 cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata
      181 gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc
      241 cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac
      301 ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg
      361 cagtacatct acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc
      421 aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc
      481 aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc
      541 gccccattga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct
      601 caataaaaga gcccacaacc cctcactcgg ggcgccagtc ctccgattga ctgagtcgcc
      661 cgggtacccg tattcccaat aaagcctctt gctgtttgca tccgaatcgt ggtctcgctg
      721 ttccttggga gggtctcctc tgagtgattg actacccacg acgggggtct ttcatttggg
      781 ggctcgtccg ggatttggag acccctgccc agggaccacc gacccaccac cgggaggtaa
      841 gctggccagc aacttatctg tgtctgtccg attgtctagt gtctatgttt gatgttatgc
      901 gcctgcgtct gtactagtta gctaactagc tctgtatctg gcggacccgt ggtggaactg
      961 acgagttctg aacacccggc cgcaaccctg ggagacgtcc cagggacttt gggggccgtt
     1021 tttgtggccc gacctgagga agggagtcga tgtggaatcc gaccccgtca ggatatgtgg
     1081 ttctggtagg agacgagaac ctaaaacagt tcccgcctcc gtctgaattt ttgctttcgg
     1141 tttggaaccg aagccgcgcg tcttgtctgc tgcagcgctg cagcatcgtt ctgtgttgtc
     1201 tctgtctgac tgtgtttctg tatttgtctg aaaattaggg ccagactgtt accactccct
     1261 taagtttgac cttaggtcac tggaaagatg tcgagcggat cgctcacaac cagtcggtag
     1321 atgtcaagaa gagacgttgg gttaccttct gctctgcaga atggccaacc tttaacgtcg
     1381 gatggccgcg agacggcacc tttaaccgag acctcatcac ccaggttaag atcaaggtct
     1441 tttcacctgg cccgcatgga cacccagacc aggtccccta catcgtgacc tgggaagcct
     1501 tggcttttga cccccctccc tgggtcaagc cctttgtaca ccctaagcct ccgcctcctc
     1561 ttcctccatc cgccccgtct ctcccccttg aacctcctcg ttcgaccccg cctcgatcct
     1621 ccctttatcc agccctcact ccttctctag gcgccggaat tcgatgtcga cattgattat
     1681 tgactagtta ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt
     1741 tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc
     1801 cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac
     1861 gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata
     1921 tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc
     1981 agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta
     2041 ttaccatggt cgaggtgagc cccacgttct gcttcactct ccccatctcc cccccctccc
     2101 cacccccaat tttgtattta tttatttttt aattattttg tgcagcgatg ggggcggggg
     2161 gggggggggg ggcgcgcgcc aggcggggcg gggcggggcg aggggcgggg cggggcgagg
     2221 cggagaggtg cggcggcagc caatcagagc ggcgcgctcc gaaagtttcc ttttatggcg
     2281 aggcggcggc ggcggcggcc ctataaaaag cgaagcgcgc ggcgggcggg agtcgctgcg
     2341 cgctgccttc gccccgtgcc ccgctccgcc gccgcctcgc gccgcccgcc ccggctctga
     2401 ctgaccgcgt tactcccaca ggtgagcggg cgggacggcc cttctcctcc gggctgtaat
     2461 tagcgcttgg tttaatgacg gcttgtttct tttctgtggc tgcgtgaaag ccttgagggg
     2521 ctccgggagg gccctttgtg cggggggagc ggctcggggg gtgcgtgcgt gtgtgtgtgc
     2581 gtggggagcg ccgcgtgcgg ctccgcgctg cccggcggct gtgagcgctg cgggcgcggc
     2641 gcggggcttt gtgcgctccg cagtgtgcgc gaggggagcg cggccggggg cggtgccccg
     2701 cggtgcgggg ggggctgcga ggggaacaaa ggctgcgtgc ggggtgtgtg cgtggggggg
     2761 tgagcagggg gtgtgggcgc gtcggtcggg ctgcaacccc ccctgcaccc ccctccccga
     2821 gttgctgagc acggcccggc ttcgggtgcg gggctccgta cggggcgtgg cgcggggctc
     2881 gccgtgccgg gcggggggtg gcggcaggtg ggggtgccgg gcggggcggg gccgcctcgg
     2941 gccggggagg gctcggggga ggggcgcggc ggcccccgga gcgccggcgg ctgtcgaggc
     3001 gcggcgagcc gcagccattg ccttttatgg taatcgtgcg agagggcgca gggacttcct
     3061 ttgtcccaaa tctgtgcgga gccgaaatct gggaggcgcc gccgcacccc ctctagcggg
     3121 cgcggggcga agcggtgcgg cgccggcagg aaggaaatgg gcggggaggg ccttcgtgcg
     3181 tcgccgcgcc gccgtcccct tctccctctc cagcctcggg gctgtccgcg gggggacggc
     3241 tgccttcggg ggggacgggg cagggcgggg ttcggcttct ggcgtgtgac cggcggctct
     3301 agagcctctg ctaaccatgt tcatgccttc ttctttttcc tacagctcct gggcaacgtg
     3361 ctggttattg tgctgtctca tcattttggc aaagaattcg ctagcggatc cggccgcctc
     3421 ggccaccggt cgccaccatg aactttctgc tctcttgggt gcactggacc ctggctttac
     3481 tgctgtacct ccaccatgcc aagtggtccc aggctgcacc cacgacagaa ggagagcaga
     3541 agtcccatga agtgatcaag ttcatggatg tctaccagcg aagctactgc cgtccgattg
     3601 agaccctggt ggacatcttc caggagtacc ccgacgagat agagtacatc ttcaagccgt
     3661 cctgtgtgcc gctgatgcgc tgtgcaggct gctgtaacga tgaagccctg gagtgcgtgc
     3721 ccacgtcaga gagcaacatc accatgcaga tcatgcggat caaacctcac caaagccagc
     3781 acataggaga gatgagcttc ctacagcaca gcagatgtga atgcagacca aagaaagaca
     3841 gaacaaagcc agaaaaatgt gacaagccaa ggcgggaagg gagaggaagc ctcctaacat
     3901 gcggtgacgt ggaggagaac ccaggaccaa tggtgagcaa gggcgaggag gataacatgg
     3961 ccatcatcaa ggagttcatg cgcttcaagg tgcacatgga gggctccgtg aacggccacg
     4021 agttcgagat cgagggcgag ggcgagggcc gcccctacga gggcacccag accgccaagc
     4081 tgaaggtgac caagggtggc cccctgccct tcgcctggga catcctgtcc cctcagttca
     4141 tgtacggctc caaggcctac gtgaagcacc ccgccgacat ccccgactac ttgaagctgt
     4201 ccttccccga gggcttcaag tgggagcgcg tgatgaactt cgaggacggc ggcgtggtga
     4261 ccgtgaccca ggactcctcc ctgcaggacg gcgagttcat ctacaaggtg aagctgcgcg
     4321 gcaccaactt cccctccgac ggccccgtaa tgcagaagaa gaccatgggc tgggaggcct
     4381 cctccgagcg gatgtacccc gaggacggcg ccctgaaggg cgagatcaag cagaggctga
     4441 agctgaagga cggcggccac tacgacgctg aggtcaagac cacctacaag gccaagaagc
     4501 ccgtgcagct gcccggcgcc tacaacgtca acatcaagtt ggacatcacc tcccacaacg
     4561 aggactacac catcgtggaa cagtacgaac gcgccgaggg ccgccactcc accggcggca
     4621 tggacgagct gtacaaggaa caaaaactta tttctgaaga agatctgtaa gtttaaacca
     4681 atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc
     4741 cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta
     4801 tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt
     4861 ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg
     4921 gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta
     4981 ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt
     5041 tgggcactga caattccgtg gtgttgtcgg ggaagctgac gtcctttcca tggctgctcg
     5101 cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca
     5161 atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc
     5221 gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcct ggaattcgag
     5281 ctcgagcttg ttaacatcga taaaataaaa gattttattt agtctccaga aaaagggggg
     5341 aatgaaagac cccacctgta ggtttggcaa gctagcttaa gtaacgccat tttgcaaggc
     5401 atggaaaaat acataactga gaatagagaa gttcagatca aggtcaggaa cagatggaac
     5461 agctgaatat gggccaaaca ggatatctgt ggtaagcagt tcctgccccg gctcagggcc
     5521 aagaacagat ggaacagctg aatatgggcc aaacaggata tctgtggtaa gcagttcctg
     5581 ccccggctca gggccaagaa cagatggtcc ccagatgcgg tccagccctc agcagtttct
     5641 agagaaccat cagatgtttc cagggtgccc caaggacctg aaatgaccct gtgccttatt
     5701 tgaactaacc aatcagttcg cttctcgctt ctgttcgcgc gcttctgctc cccgagctca
     5761 ataaaagagc ccacaacccc tcactcgggg cgccagtcct ccgattgact gagtcgcccg
     5821 ggtacccgtg tatccaataa accctcttgc agttgcatcc gacttgtggt ctcgctgttc
     5881 cttgggaggg tctcctctga gtgattgact acccgtcagc gggggtcttt cacatgcagc
     5941 atgtatcaaa attaatttgg ttttttttct taagtattta cattaaatgg ccatagttgc
     6001 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     6061 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     6121 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     6181 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     6241 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     6301 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     6361 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     6421 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     6481 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     6541 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     6601 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     6661 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     6721 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     6781 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     6841 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     6901 tatcaaaaag gatcttcacc tagatccttt tgcggccgca aatcaatcta aagtatatat
     6961 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     7021 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     7081 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     7141 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     7201 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     7261 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     7321 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     7381 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     7441 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     7501 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     7561 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     7621 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     7681 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     7741 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     7801 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatga
     7861 tccggccatt agccatatta ttcattggtt atatagcata aatcaatatt ggctattggc
     7921 cattgcatac gttgtatcca tatcataata tgtacattta tattggctca tgtcca
//
