LOCUS       sequence-405333-        7786 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Conditional expression of target gene with fluorescent reporter 
            mScarlet.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7786)
  AUTHORS   Fisher J, Verhagen M, Long Z, Moissidis M, Yan Y, He C, Wang J, 
            Micoli E, Alastruey CM, Moors R, Marin O, Mi D, Lim L
  TITLE     Cortical somatostatin long-range projection neurons and interneurons
            exhibit divergent developmental trajectories.
  JOURNAL   Neuron. 2023 Dec 7:S0896-6273(23)00887-5. doi: 
            10.1016/j.neuron.2023.11.013.
  PUBMED    38086373
REFERENCE   2  (bases 1 to 7786)
  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: "Neuron. 
            2023 Dec 7:S0896-6273(23)00887-5. doi: 
            10.1016/j.neuron.2023.11.013."
COMMENT     Sequence Label: p185-RV-CAG-dio-mScarlet-T2A
FEATURES             Location/Qualifiers
     source          1..7786
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             182..651
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(781..798)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(952..1540)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1032..1051)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1700..2491,2492..2560))
                     /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     2323..2342
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     enhancer        2712..3091
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3092..3295
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3246..3266
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             3296..3471
                     /label=5' LTR (truncated)
                     /note="truncated long terminal repeat from Moloney murine 
                     sarcoma virus"
     misc_feature    3534..3733
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             3934..4350
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     4280..4302
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     4318..4334
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     enhancer        4364..4743
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4745..5021
                     /label=chicken beta-actin promoter
     intron          5022..6038
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     5962..5984
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     6046..6065
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     protein_bind    complement(6108..6141)
                     /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(6192..6225)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(6246..6299)
                     /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(6300..6995)
                     /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(6575..6594)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     6844..6862
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     protein_bind    7070..7103
                     /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    7154..7187
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    7197..7785
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(7250..7270)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(7668..7679)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
ORIGIN
        1 tggaattcga gctcgacgat aaaataaaag attttattta gtctccagaa aaagggggga
       61 atgaaagacc ccacctgtag gtttggcaag ctagcttaag taacgccatt ttgcaaggca
      121 tggaaaaata cataactgag aatagagaag ttcagatcaa ggtcaggaac agatggaaca
      181 gctgaatatg ggccaaacag gatatctgtg gtaagcagtt cctgccccgg ctcagggcca
      241 agaacagatg gaacagctga atatgggcca aacaggatat ctgtggtaag cagttcctgc
      301 cccggctcag ggccaagaac agatggtccc cagatgcggt ccagccctca gcagtttcta
      361 gagaaccatc agatgtttcc agggtgcccc aaggacctga aatgaccctg tgccttattt
      421 gaactaacca atcagttcgc ttctcgcttc tgttcgcgcg cttctgctcc ccgagctcaa
      481 taaaagagcc cacaacccct cactcggggc gccagtcctc cgattgactg agtcgcccgg
      541 gtacccgtgt atccaataaa ccctcttgca gttgcatccg acttgtggtc tcgctgttcc
      601 ttgggagggt ctcctctgag tgattgacta cccgtcagcg ggggtctttc acatgcagca
      661 tgtatcaaaa ttaatttggt tttttttctt aagtatttac attaaatggc catagttgca
      721 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
      781 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
      841 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
      901 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
      961 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     1021 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     1081 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     1141 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     1201 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     1261 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     1321 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     1381 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     1441 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     1501 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     1561 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     1621 atcaaaaagg atcttcacct agatcctttt gcggccgcaa atcaatctaa agtatatatg
     1681 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     1741 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     1801 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     1861 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     1921 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     1981 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     2041 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     2101 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     2161 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     2221 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     2281 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     2341 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     2401 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     2461 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     2521 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatgat
     2581 ccggccatta gccatattat tcattggtta tatagcataa atcaatattg gctattggcc
     2641 attgcatacg ttgtatccat atcataatat gtacatttat attggctcat gtccaacatt
     2701 accgccatgt tgacattgat tattgactag ttattaatag taatcaatta cggggtcatt
     2761 agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg
     2821 ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac
     2881 gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt
     2941 ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa
     3001 atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta
     3061 catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt acatcaatgg
     3121 gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg
     3181 gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc
     3241 attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca gagctcaata
     3301 aaagagccca caacccctca ctcggggcgc cagtcctccg attgactgag tcgcccgggt
     3361 acccgtattc ccaataaagc ctcttgctgt ttgcatccga atcgtggtct cgctgttcct
     3421 tgggagggtc tcctctgagt gattgactac ccacgacggg ggtctttcat ttgggggctc
     3481 gtccgggatt tggagacccc tgcccaggga ccaccgaccc accaccggga ggtaagctgg
     3541 ccagcaactt atctgtgtct gtccgattgt ctagtgtcta tgtttgatgt tatgcgcctg
     3601 cgtctgtact agttagctaa ctagctctgt atctggcgga cccgtggtgg aactgacgag
     3661 ttctgaacac ccggccgcaa ccctgggaga cgtcccaggg actttggggg ccgtttttgt
     3721 ggcccgacct gaggaaggga gtcgatgtgg aatccgaccc cgtcaggata tgtggttctg
     3781 gtaggagacg agaacctaaa acagttcccg cctccgtctg aatttttgct ttcggtttgg
     3841 aaccgaagcc gcgcgtcttg tctgctgcag cgctgcagca tcgttctgtg ttgtctctgt
     3901 ctgactgtgt ttctgtattt gtctgaaaat tagggccaga ctgttaccac tcccttaagt
     3961 ttgaccttag gtcactggaa agatgtcgag cggatcgctc acaaccagtc ggtagatgtc
     4021 aagaagagac gttgggttac cttctgctct gcagaatggc caacctttaa cgtcggatgg
     4081 ccgcgagacg gcacctttaa ccgagacctc atcacccagg ttaagatcaa ggtcttttca
     4141 cctggcccgc atggacaccc agaccaggtc ccctacatcg tgacctggga agccttggct
     4201 tttgaccccc ctccctgggt caagcccttt gtacacccta agcctccgcc tcctcttcct
     4261 ccatccgccc cgtctctccc ccttgaacct cctcgttcga ccccgcctcg atcctccctt
     4321 tatccagccc tcactccttc tctaggcgcc ggaattcgat gtcgacattg attattgact
     4381 agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc
     4441 gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg
     4501 acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa
     4561 tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca
     4621 agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac
     4681 atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc
     4741 atggtcgagg tgagccccac gttctgcttc actctcccca tctccccccc ctccccaccc
     4801 ccaattttgt atttatttat tttttaatta ttttgtgcag cgatgggggc gggggggggg
     4861 gggggcgcgc gccaggcggg gcggggcggg gcgaggggcg gggcggggcg aggcggagag
     4921 gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt tccttttatg gcgaggcggc
     4981 ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc gggagtcgct gcgcgctgcc
     5041 ttcgccccgt gccccgctcc gccgccgcct cgcgccgccc gccccggctc tgactgaccg
     5101 cgttactccc acaggtgagc gggcgggacg gcccttctcc tccgggctgt aattagcgct
     5161 tggtttaatg acggcttgtt tcttttctgt ggctgcgtga aagccttgag gggctccggg
     5221 agggcccttt gtgcgggggg agcggctcgg ggggtgcgtg cgtgtgtgtg tgcgtgggga
     5281 gcgccgcgtg cggctccgcg ctgcccggcg gctgtgagcg ctgcgggcgc ggcgcggggc
     5341 tttgtgcgct ccgcagtgtg cgcgagggga gcgcggccgg gggcggtgcc ccgcggtgcg
     5401 gggggggctg cgaggggaac aaaggctgcg tgcggggtgt gtgcgtgggg gggtgagcag
     5461 ggggtgtggg cgcgtcggtc gggctgcaac cccccctgca cccccctccc cgagttgctg
     5521 agcacggccc ggcttcgggt gcggggctcc gtacggggcg tggcgcgggg ctcgccgtgc
     5581 cgggcggggg gtggcggcag gtgggggtgc cgggcggggc ggggccgcct cgggccgggg
     5641 agggctcggg ggaggggcgc ggcggccccc ggagcgccgg cggctgtcga ggcgcggcga
     5701 gccgcagcca ttgcctttta tggtaatcgt gcgagagggc gcagggactt cctttgtccc
     5761 aaatctgtgc ggagccgaaa tctgggaggc gccgccgcac cccctctagc gggcgcgggg
     5821 cgaagcggtg cggcgccggc aggaaggaaa tgggcgggga gggccttcgt gcgtcgccgc
     5881 gccgccgtcc ccttctccct ctccagcctc ggggctgtcc gcggggggac ggctgccttc
     5941 gggggggacg gggcagggcg gggttcggct tctggcgtgt gaccggcggc tctagagcct
     6001 ctgctaacca tgttcatgcc ttcttctttt tcctacagct cctgggcaac gtgctggtta
     6061 ttgtgctgtc tcatcatttt ggcaaagaat tcgctagcgg atccggaata acttcgtata
     6121 ggatacttta tacgaagtta tgcagaatgg tagctggatt gtagctgcta ttagcaatat
     6181 gaaacctctt aataacttcg tatagcatac attatacgaa gttatttaat taaatgatgc
     6241 tcgagagggc cgggattctc ttccacgtct ccgcatgtca gaagacttcc tctgccctcc
     6301 ttgtacagct cgtccatgcc gccggtggag tggcggccct cggagcgttc gtactgttcc
     6361 accacggtgt agtcctcgtt gtgggaggtg atgtccaact tgcggtcgac gttgtaggcg
     6421 ccgggcatct gcacgggctt cttggccttg taggtggtct tgaagtccgc caggtagcgg
     6481 ccgccgtcct tcaggcgcag ggccatctta atgtcgccct tcagcacgcc gtcctcgggg
     6541 tacaaccgct cggtggacgc ttcccagccc attgtcttct tctgcattac ggggccgtca
     6601 ggagggaagt tggtgccgcg gagcttcacc ttgtagatca gggtgccgtc ctccagggag
     6661 gtgtcctggg tcacggtcac ggcgccgccg tcctcgaagt tcatcacgcg ctcccacttg
     6721 aagccctcgg ggaaggactg cttatagtag tcggggatgt cggcggggtg cttgatgaag
     6781 gccctggagc cgtacatgaa ctgaggggac aggatgtccc aggagaaggg cagggggcca
     6841 cccttggtca ccttcagctt ggcggtctgg gtgccctcgt aggggcggcc ctcgccctcg
     6901 ccctcgatct cgaactcgtg gccgttcatg gagccctcca tgtgcacctt gaaccgcatg
     6961 aactccttga tcactgcctc gcccttgctc accatgatct tttggtcctc atcattcttt
     7021 tcaacctgtt tggttcttct catacagcac agcatggtgg cgcgccgcga taacttcgta
     7081 taaagtatcc tatacgaagt tatttgcctt aacccagaaa ttatcactgt tattctttag
     7141 aatggtgcaa agaataactt cgtataatgt atgctatacg aagttatgtt taaaccaatc
     7201 aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt
     7261 ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg
     7321 ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc
     7381 ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt
     7441 ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg
     7501 ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg
     7561 gcactgacaa ttccgtggtg ttgtcgggga agctgacgtc ctttccatgg ctgctcgcct
     7621 gtgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc
     7681 cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc
     7741 ttcgccctca gacgagtcgg atctcccttt gggccgcctc cccgcc
//
