LOCUS       sequence-461407-        8378 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For strong lentivirus-mediated expression of dTomato.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8378)
  AUTHORS   Shin D, Kim CN, Ross J, Hennick KM, Wu SR, Paranjape N, Leonard R, 
            Wang JC, Keefe MG, Pavlovic BJ, Donohue KC, Moreau C, Wigdor EM, 
            Larson HH, Allen DE, Cadwell CR, Bhaduri A, Popova G, Bearden CE, 
            Pollen AA, Jacquemont S, Sanders SJ, Haussler D, Wiita AP, Frost NA,
            Sohal VS, Nowakowski TJ
  TITLE     Thalamocortical organoids enable in vitro modeling of 22q11.2 
            microdeletion associated with neuropsychiatric disorders.
  JOURNAL   Cell Stem Cell. 2024 Mar 7;31(3):421-432.e8. doi: 
            10.1016/j.stem.2024.01.010. Epub 2024 Feb 20.
  PUBMED    38382530
REFERENCE   2  (bases 1 to 8378)
  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"; doi: 
            "10.1016/j.stem.2024.01.010"; journalName: "Cell Stem Cell"; date: 
            "2024-03-7- 7"; volume: "31"; issue: "3"; pages: "421-432.e8"
COMMENT     Sequence Label: pSicoFlipped-CAG-dTomato-WPRE-bGHpA
FEATURES             Location/Qualifiers
     source          1..8378
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             337..381
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    679..796
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     polyA_signal    854..1078
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     misc_feature    1180..1768
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             1286..1297
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     1695..1715
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(1805..2506)
                     /codon_start=1
                     /product="dimeric variant of DsRed fluorescent protein 
                     (Shaner et al., 2004)"
                     /label=dTomato
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     LVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH
                     HLFLYGMDELYK"
     primer_bind     complement(1888..1911)
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     primer_bind     complement(2086..2105)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     2355..2373
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     intron          2537..3553
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     complement(2591..2613)
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     promoter        complement(3554..3829)
                     /label=chicken beta-actin promoter
     enhancer        3831..4210
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     LTR             4619..4799
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(4861..5449)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4941..4960)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5620..6411,6412..6480))
                     /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     6243..6262
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6481..6585)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6660..6679)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        6851..7230
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7232..7430
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7385..7405
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             7448..7628
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    7675..7800
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    join(8297..8378,1..152)
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
ORIGIN
        1 caattattgt ctggtatagt gcagcagcag aacaatttgc tgagggctat tgaggcgcaa
       61 cagcatctgt tgcaactcac agtctggggc atcaagcagc tccaggcaag aatcctggct
      121 gtggaaagat acctaaagga tcaacagctc ctggggattt ggggttgctc tggaaaactc
      181 atttgcacca ctgctgtgcc ttggaatgct agttggagta ataaatctct ggaacagatt
      241 tggaatcaca cgacctggat ggagtgggac agagaaatta acaattacac aagcttaata
      301 cactccttaa ttgaagaatc gcaaaaccag caagaaaaga atgaacaaga attattggaa
      361 ttagataaat gggcaagttt gtggaattgg tttaacataa caaattggct gtggtatata
      421 aaattattca taatgatagt aggaggcttg gtaggtttaa gaatagtttt tgctgtactt
      481 tctatagtga atagagttag gcagggatat tcaccattat cgtttcagac ccacctccca
      541 accccgaggg gacccgacag gcccgaagga atagaagaag aaggtggaga gagagacaga
      601 gacagatcca ttcgattagt gaacggatcg gcactgcgtg cgccaattct gcagacaaat
      661 ggcagtattc atccacaatt ttaaaagaaa aggggggatt ggggggtaca gtgcagggga
      721 aagaatagta gacataatag caacagacat acaaactaaa gaattacaaa aacaaattac
      781 aaaaattcaa aattttcggg tttattacag ggacagcaga gatccagttt ggttagtacc
      841 gggcccgctc tagccataga gcccaccgca tccccagcat gcctgctatt gtcttcccaa
      901 tcctccccct tgctgtcctg ccccacccca ccccccagaa tagaatgaca cctactcaga
      961 caatgcgatg caatttcctc attttattag gaaaggacag tgggagtggc accttccagg
     1021 gtcaaggaag gcacggggga ggggcaaaca acagatggct ggcaactaga aggcacagct
     1081 cgaggccggt gtaatcagcc tagctacatg ccccatgacg atccacggat gtgctgtctg
     1141 aattcgcttg tgccccagtt tgctagggag gtccaggacg cggggaggcg gcccaaaggg
     1201 agatccgact cgtctgaggg cgaaggcgaa gacgcggaag aggccgcaga gccggcagca
     1261 ggccgcggga aggaaggtcc gctggattga gggccgaagg gacgtagcag aaggacgtcc
     1321 cgcgcagaat ccaggtggca acacaggcga gcagccaagg aaaggacgat gatttccccg
     1381 acaacaccac ggaattgtca gtgcccaaca gccgagcccc tgtccagcag cgggcaaggc
     1441 aggcggcgat gagttccgcc gtggcaatag ggagggggaa agcgaaagtc ccggaaagga
     1501 gctgacaggt ggtggcaatg ccccaaccag tgggggttgc gtcagcaaac acagtgcaca
     1561 ccacgccacg ttgcctgaca acgggccaca actcctcata aagagacagc aaccaggatt
     1621 tatacaagga ggagaaaatg aaagccatac gggaagcaat agcatgatac aaaggcatta
     1681 aagcagcgta tccacatagc gtaaaaggag caacatagtt aagaatacca gtcaatcttt
     1741 cacaaatttt gtaatccaga ggttgattgg atcgtacgtc ggcataagct ttgtagcggc
     1801 cctacttgta cagctcgtcc atgccgtaca ggaacaggtg gtggcggccc tcggagcgct
     1861 cgtactgttc cacgatggtg tagtcctcgt tgtgggaggt gatgtccagc ttggtgtcca
     1921 cgtagtagta gccgggcagt tgcacgggct tcttggccat gtagatggtc ttgaactcca
     1981 ccaggtagtg gccgccgtcc ttcagcttca gggcctggtg gatctcgccc ttcagcacgc
     2041 cgtcgcgggg gtacaggcgc tcggtggagg cctcccagcc catggtcttc ttctgcatta
     2101 cggggccgtc gggggggaag ttggtgccgc gcatcttcac cttgtagatc agcgtgccgt
     2161 cctgcaggga ggagtcctgg gtcacggtca ccagaccgcc gtcctcgaag ttcatcacgc
     2221 gctcccactt gaagccctcg gggaaggaca gcttcttgta atcggggatg tcggcggggt
     2281 gcttcacgta cgccttggag ccgtacatga actgggggga caggatgtcc caggcgaagg
     2341 gcagggggcc gcccttggtc accttcagct tggcggtctg ggtgccctcg taggggcggc
     2401 cctcgccctc gccctcgatc tcgaactcgt ggccgttcat ggagccctcc atgcgcacct
     2461 tgaagcgcat gaactctttg atgacctcct cgcccttgct caccataccg gtagcgctag
     2521 ttacgactag caggagctgt aggaaaaaga agaaggcatg aacatggtta gcagaggctc
     2581 tagagccgcc ggtcacacgc cagaagccga accccgccct gccccgtccc ccccgaaggc
     2641 agccgtcccc ccgcggacag ccccgaggct ggagagggag aaggggacgg cggcgcggcg
     2701 acgcacgaag gccctccccg cccatttcct tcctgccggc gccgcaccgc ttcgccccgc
     2761 gcccgctaga gggggtgcgg cggcgcctcc cagatttcgg ctccgcacag atttgggaca
     2821 aaggaagtcc ctgcgccctc tcgcacgatt accataaaag gcaatggctg cggctcgccg
     2881 cgcctcgaca gccgccggcg ctccgggggc cgccgcgccc ctcccccgag ccccccccgg
     2941 cccgaggcgg ccccgccccg cccggcaccc ccacctgccg ccaccccccg cccggcacgg
     3001 cgagccccgc gccacgcccc gtacggagcc ccgcacccga agccgggccg tgctcagcaa
     3061 ctcggggagg ggggtgcagg gggggttgca gcccgaccga cgcgcccaca ccccctgctc
     3121 acccccccac gcacacaccc cgcacgcagc ctttgttccc ctcgcagccc cccccgcacc
     3181 gcggggcacc gcccccggcc gcgctcccct cgcgcacact gcggagcgca caaagccccg
     3241 cgccgcgccc gcagcgctca cagccgccgg gcagcgcgga gccgcacgcg gcgctcccca
     3301 cgcacacaca cacgcacgca ccccccgagc cgctcccccc gcacaaaggg ccctcccgga
     3361 gcccctcaag gctttcacgc agccacagaa aagaaacaag ccgtcattaa accaagcgct
     3421 aattacagcc cggaggagaa gggccgtccc gcccgctcac ctgtgggagt aacgcggtca
     3481 gtcagagccg gggcgggcgg cgcgaggcgg cggcggagcg gggcacgggg cgaaggcagc
     3541 gcgcagcgac tcccgcccgc cgcgcgcttc gctttttata gggccgccgc cgccgccgcc
     3601 tcgccataaa aggaaacttt cggagcgcgc cgctctgatt ggctgccgcc gcacctctcc
     3661 gcctcgcccc cccccccccc cccccccccc cccccccccc cggcgcgccc cccccccccc
     3721 cccgccccca tcgctgcaca aaataattaa aaaataaata aatacaaaat tgggggtggg
     3781 gagggggggg agatggggag agtgaagcag aacgtggggc tcacctcgac catggtaata
     3841 gcgatgacta atacgtagat gtactgccaa gtaggaaagt cccataaggt catgtactgg
     3901 gcataatgcc aggcgggcca tttaccgtca ttgacgtcaa tagggggcgt acttggcata
     3961 tgatacactt gatgtactgc caagtgggca gtttaccgta aatactccac ccattgacgt
     4021 caatggaaag tccctattgg cgttactatg ggaacatacg tcattattga cgtcaatggg
     4081 cgggggtcgt tgggcggtca gccaggcggg ccatttaccg taagttatgt aacgcggaac
     4141 tccatatatg ggctatgaac taatgacccc gtaattgatt actattaata actagtcaat
     4201 aatcaatgtc gagctgatct acgatgtagt acctttaaga ccaatgactt acaaggcagc
     4261 tgtagatctt agccactttt taaaagaaaa ggggggactg gaagggctaa ttcactccca
     4321 acgaagacaa gatatccttg atctgtggat ctaccacaca caaggctact tccctgattg
     4381 gcagaactac acaccagggc cagggatcag atatccactg acctttggat ggtgctacaa
     4441 gctagtacca gttgagcaag agaaggtaga agaagccaat gaaggagaga acacccgctt
     4501 gttacaccct gtgagcctgc atgggatgga tgacccggag agagaagtat tagagtggag
     4561 gtttgacagc cgcctagcat ttcatcacat ggcccgagag ctgcatccgg actgtactgg
     4621 gtctctctgg ttagaccaga tctgagcctg ggagctctct ggctaactag ggaacccact
     4681 gcttaagcct caataaagct tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg
     4741 tgactctggt aactagagat ccctcagacc cttttagtca gtgtggaaaa tctctagcag
     4801 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     4861 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     4921 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     4981 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     5041 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     5101 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     5161 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     5221 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     5281 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     5341 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     5401 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     5461 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     5521 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     5581 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     5641 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     5701 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     5761 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     5821 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     5881 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     5941 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     6001 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     6061 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     6121 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     6181 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     6241 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     6301 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     6361 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     6421 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     6481 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     6541 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     6601 agtgccacct gacgtcgacg gatcgggaga tctcccgatc ccctatggtg cactctcagt
     6661 acaatctgct ctgatgccgc atagttaagc cagtatctgc tccctgcttg tgtgttggag
     6721 gtcgctgagt agtgcgcgag caaaatttaa gctacaacaa ggcaaggctt gaccgacaat
     6781 tgcatgaaga atctgcttag ggttaggcgt tttgcgctgc ttcgcgatgt acgggccaga
     6841 tatacgcgtt gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta
     6901 gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc
     6961 tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg
     7021 ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg
     7081 gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa
     7141 tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac
     7201 atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg
     7261 cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg
     7321 agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca
     7381 ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag cgcgttttgc
     7441 ctgtactggg tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg
     7501 gaacccactg cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg
     7561 tctgttgtgt gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat
     7621 ctctagcagt ggcgcccgaa cagggacttg aaagcgaaag ggaaaccaga ggagctctct
     7681 cgacgcagga ctcggcttgc tgaagcgcgc acggcaagag gcgaggggcg gcgactggtg
     7741 agtacgccaa aaattttgac tagcggaggc tagaaggaga gagatgggtg cgagagcgtc
     7801 agtattaagc gggggagaat tagatcgcga tgggaaaaaa ttcggttaag gccaggggga
     7861 aagaaaaaat ataaattaaa acatatagta tgggcaagca gggagctaga acgattcgca
     7921 gttaatcctg gcctgttaga aacatcagaa ggctgtagac aaatactggg acagctacaa
     7981 ccatcccttc agacaggatc agaagaactt agatcattat ataatacagt agcaaccctc
     8041 tattgtgtgc atcaaaggat agagataaaa gacaccaagg aagctttaga caagatagag
     8101 gaagagcaaa acaaaagtaa gaccaccgca cagcaagcgg ccggccgctg atcttcagac
     8161 ctggaggagg agatatgagg gacaattgga gaagtgaatt atataaatat aaagtagtaa
     8221 aaattgaacc attaggagta gcacccacca aggcaaagag aagagtggtg cagagagaaa
     8281 aaagagcagt gggaatagga gctttgttcc ttgggttctt gggagcagca ggaagcacta
     8341 tgggcgcagc gtcaatgacg ctgacggtac aggccaga
//
