LOCUS       sequence-335020-        8405 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral vector expressing human LMX1A (with silent mutation) with
            copGFP reporter gene, under control of EF1alpha promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8405)
  AUTHORS   Wulansari N, Darsono WHW, Woo HJ, Chang MY, Kim J, Bae EJ, Sun W, 
            Lee JH, Cho IJ, Shin H, Lee SJ, Lee SH
  TITLE     Neurodevelopmental defects and neurodegenerative phenotypes in human
            brain organoids carrying Parkinson's disease-linked DNAJC6 
            mutations.
  JOURNAL   Sci Adv. 2021 Feb 17;7(8). pii: 7/8/eabb1540. doi: 
            10.1126/sciadv.abb1540. Print 2021 Feb.
  PUBMED    33597231
REFERENCE   2  (bases 1 to 8405)
  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: "Sci Adv."; 
            date: "2021-02-17"; volume: "7(8). pii: 7/8/eabb1540. doi"; pages: "
            10.1126/sciadv.abb1540. Print 2021 Feb"
COMMENT     Sequence Label: pCDH-EF1a-hLMX1A-T2A-copGFP
FEATURES             Location/Qualifiers
     source          1..8405
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        6..232
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             233..413
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    460..585
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1078..1311
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1496..1540
                     /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    1807..1923
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        1966..2177
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     LTR             2190..2458
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     CDS             3647..3700
                     /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             3731..4393
                     /codon_start=1
                     /product="green fluorescent protein 2 from Pontellina 
                     plumata, also known as ppluGFP2 (Shagin et al., 2004)"
                     /label=CopGFP
                     /translation="PAMEIECRITGTLNGVEFELVGGGEGTPKQGRMTNKMKSTKGALT
                     FSPYLLSHVMGYGFYHFGTYPSGYENPFLHAINNGGYTNTRIEKYEDGGVLHVSFSYRY
                     EAGRVIGDFKVVGTGFPEDSVIFTDKIIRSNATVEHLHPMGDNVLVGSFARTFSLRDGG
                     YYSFVVDSHMHFKSAIHPSILQNGGPMFAFRRVEELHSNTELGIVEYQHAFKTPIAFA"
     misc_feature    4484..5072
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4537..4557)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4955..4966)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             5146..5379
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    5451..5572
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5488..5507)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5542..5561
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      5612..5747
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(5780..5796)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5780..5796)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5793..5815)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5804..5820
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(5828..5834,5835..5852,5853..5858))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5873..5894
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6011..6028)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6182..6770)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(join(6941..7732,7733..7801))
                     /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     7564..7583
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7802..7906)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7974..7992
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(8030..8052)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     8152..8171
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     8365..8387
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     8379..8396
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     8380..8396
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 acgcgtgtag tcttatgcaa tactcttgta gtcttgcaac atggtaacga tgagttagca
       61 acatgcctta caaggagaga aaaagcaccg tgcatgccga ttggtggaag taaggtggta
      121 cgatcgtgcc ttattaggaa ggcaacagac gggtctgaca tggattggac gaaccactga
      181 attgccgcat tgcagagata ttgtatttaa gtgcctagct cgatacataa acgggtctct
      241 ctggttagac cagatctgag cctgggagct ctctggctaa ctagggaacc cactgcttaa
      301 gcctcaataa agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc
      361 tggtaactag agatccctca gaccctttta gtcagtgtgg aaaatctcta gcagtggcgc
      421 ccgaacaggg acttgaaagc gaaagggaaa ccagaggagc tctctcgacg caggactcgg
      481 cttgctgaag cgcgcacggc aagaggcgag gggcggcgac tggtgagtac gccaaaaatt
      541 ttgactagcg gaggctagaa ggagagagat gggtgcgaga gcgtcagtat taagcggggg
      601 agaattagat cgcgatggga aaaaattcgg ttaaggccag ggggaaagaa aaaatataaa
      661 ttaaaacata tagtatgggc aagcagggag ctagaacgat tcgcagttaa tcctggcctg
      721 ttagaaacat cagaaggctg tagacaaata ctgggacagc tacaaccatc ccttcagaca
      781 ggatcagaag aacttagatc attatataat acagtagcaa ccctctattg tgtgcatcaa
      841 aggatagaga taaaagacac caaggaagct ttagacaaga tagaggaaga gcaaaacaaa
      901 agtaagacca ccgcacagca agcggccact gatcttcaga cctggaggag gagatatgag
      961 ggacaattgg agaagtgaat tatataaata taaagtagta aaaattgaac cattaggagt
     1021 agcacccacc aaggcaaaga gaagagtggt gcagagagaa aaaagagcag tgggaatagg
     1081 agctttgttc cttgggttct tgggagcagc aggaagcact atgggcgcag cgtcaatgac
     1141 gctgacggta caggccagac aattattgtc tggtatagtg cagcagcaga acaatttgct
     1201 gagggctatt gaggcgcaac agcatctgtt gcaactcaca gtctggggca tcaagcagct
     1261 ccaggcaaga atcctggctg tggaaagata cctaaaggat caacagctcc tggggatttg
     1321 gggttgctct ggaaaactca tttgcaccac tgctgtgcct tggaatgcta gttggagtaa
     1381 taaatctctg gaacagattt ggaatcacac gacctggatg gagtgggaca gagaaattaa
     1441 caattacaca agcttaatac actccttaat tgaagaatcg caaaaccagc aagaaaagaa
     1501 tgaacaagaa ttattggaat tagataaatg ggcaagtttg tggaattggt ttaacataac
     1561 aaattggctg tggtatataa aattattcat aatgatagta ggaggcttgg taggtttaag
     1621 aatagttttt gctgtacttt ctatagtgaa tagagttagg cagggatatt caccattatc
     1681 gtttcagacc cacctcccaa ccccgagggg acccgacagg cccgaaggaa tagaagaaga
     1741 aggtggagag agagacagag acagatccat tcgattagtg aacggatctc gacggtatcg
     1801 gttaactttt aaaagaaaag gggggattgg ggggtacagt gcaggggaaa gaatagtaga
     1861 cataatagca acagacatac aaactaaaga attacaaaaa caaattacaa aattcaaaat
     1921 tttatcgata ctagtggatc tgcgatcgct ccggtgcccg tcagtgggca gagcgcacat
     1981 cgcccacagt ccccgagaag ttggggggag gggtcggcaa ttgaacgggt gcctagagaa
     2041 ggtggcgcgg ggtaaactgg gaaagtgatg tcgtgtactg gctccgcctt tttcccgagg
     2101 gtgggggaga accgtatata agtgcagtag tcgccgtgaa cgttcttttt cgcaacgggt
     2161 ttgccgccag aacacagctg aagcttcgag gggctcgcat ctctccttca cgcgcccgcc
     2221 gccctacctg aggccgccat ccacgccggt tgagtcgcgt tctgccgcct cccgcctgtg
     2281 gtgcctcctg aactgcgtcc gccgtctagg taagtttaaa gctcaggtcg agaccgggcc
     2341 tttgtccggc gctcccttgg agcctaccta gactcagccg gctctccacg ctttgcctga
     2401 ccctgcttgc tcaactctac gtctttgttt cgttttctgt tctgcgccgt tacagatcca
     2461 agctgtgacc ggcgcctact ctagagctag catgctggac ggcctaaaga tggaggagaa
     2521 cttccaaagc gcgatcgaca cctcggcctc cttctcctcg ctgctgggca gagcggtgag
     2581 ccccaagtct gtctgcgagg gctgtcagcg ggtcatcttg gacaggtttc tgctgcggct
     2641 caacgacagc ttctggcatg agcagtgcgt gcagtgcgcc tcctgcaaag agcccctgga
     2701 gaccacctgc ttctaccggg acaagaagct gtactgcaag tatgactacg agaagctgtt
     2761 tgctgttaaa tgtgggggct gcttcgaggc catcgctccc aatgagtttg ttatgcgggc
     2821 ccagaagagt gtataccacc tgagctgctt ctgttgctgt gtctgcgagc gacagcttca
     2881 gaagggtgat gagtttgtcc tgaaggaggg gcagctgctc tgcaaagggg actatgagaa
     2941 ggagcgggag ctactcagcc tggtgagccc agcagcctca gactcaggta aaagtgatga
     3001 tgaagaaagt ctctgcaagt cagcccatgg ggcagggaaa ggaactgctg aggaaggcaa
     3061 ggaccataag cgccccaaac gtccgagaac catcttgaca actcaacaga ggcgagcatt
     3121 caaggcctca tttgaagtat cctccaagcc ctgcaggaag gtgagagaga ctctggctgc
     3181 agagacaggg ctgagtgtcc gtgtcgtcca ggtgtggttc caaaaccaga gagcgaagat
     3241 gaagaagctg gccaggcgac agcagcagca gcagcaagat cagcagaaca cccagaggct
     3301 gagctctgct cagacaaacg gtggtgggag tgctgggatg gaaggaatca tgaaccccta
     3361 cacggctctg cccaccccac agcagctcct ggccatcgag cagagtgtct acagctcaga
     3421 tcccttccga cagggtctca ccccacccca gatgcctgga gaccacatgc acccttatgg
     3481 tgccgagccc cttttccatg acctggatag cgacgacacc tccctcagta acctgggtga
     3541 ctgtttccta gcaacctcag aagctgggcc tctgcagtcc agagtgggaa accccattga
     3601 ccatctgtac tccatgcaga attcttactt cacatctgcg gccgctgagg gcagaggaag
     3661 tcttctaaca tgcggtgacg tggaggagaa tcccggccct tccggaatgg agagcgacga
     3721 gagcggcctg cccgccatgg agatcgagtg ccgcatcacc ggcaccctga acggcgtgga
     3781 gttcgagctg gtgggcggcg gagagggcac ccccaagcag ggccgcatga ccaacaagat
     3841 gaagagcacc aaaggcgccc tgaccttcag cccctacctg ctgagccacg tgatgggcta
     3901 cggcttctac cacttcggca cctaccccag cggctacgag aaccccttcc tgcacgccat
     3961 caacaacggc ggctacacca acacccgcat cgagaagtac gaggacggcg gcgtgctgca
     4021 cgtgagcttc agctaccgct acgaggccgg ccgcgtgatc ggcgacttca aggtggtggg
     4081 caccggcttc cccgaggaca gcgtgatctt caccgacaag atcatccgca gcaacgccac
     4141 cgtggagcac ctgcacccca tgggcgataa cgtgctggtg ggcagcttcg cccgcacctt
     4201 cagcctgcgc gacggcggct actacagctt cgtggtggac agccacatgc acttcaagag
     4261 cgccatccac cccagcatcc tgcagaacgg gggccccatg ttcgccttcc gccgcgtgga
     4321 ggagctgcac agcaacaccg agctgggcat cgtggagtac cagcacgcct tcaagacccc
     4381 catcgccttc gccagatccc gcgctcagtc gtccaattct gccgtggacg gcaccgccgg
     4441 acccggctcc accggatctc gctagagctg aatctaagtc gacaatcaac ctctggatta
     4501 caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg
     4561 atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc
     4621 ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca
     4681 acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac
     4741 cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact
     4801 catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc
     4861 cgtggtgttg tcggggaaat catcgtcctt tccttggctg ctcgcctgtg ttgccacctg
     4921 gattctgcgc gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc
     4981 ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac
     5041 gagtcggatc tccctttggg ccgcctcccc gcctggtacc tttaagacca atgacttaca
     5101 aggcagctgt agatcttagc cactttttaa aagaaaaggg gggactggaa gggctaattc
     5161 actcccaacg aagataagat ctgctttttg cttgtactgg gtctctctgg ttagaccaga
     5221 tctgagcctg ggagctctct ggctaactag ggaacccact gcttaagcct caataaagct
     5281 tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt aactagagat
     5341 ccctcagacc cttttagtca gtgtggaaaa tctctagcag tagtagttca tgtcatctta
     5401 ttattcagta tttataactt gcaaagaaat gaatatcaga gagtgagagg aacttgttta
     5461 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     5521 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct
     5581 ggctctagct atcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc
     5641 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggcag aggccgcctc
     5701 ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag acttttgcag
     5761 agaccaaatt cgtaatcatg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
     5821 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
     5881 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     5941 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     6001 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     6061 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     6121 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     6181 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     6241 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     6301 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     6361 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     6421 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     6481 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     6541 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     6601 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     6661 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     6721 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     6781 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     6841 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     6901 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     6961 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     7021 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     7081 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     7141 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     7201 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     7261 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     7321 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     7381 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     7441 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     7501 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     7561 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     7621 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     7681 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     7741 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     7801 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     7861 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     7921 aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc
     7981 gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca
     8041 tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc
     8101 gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag
     8161 agcagattgt actgagagtg caccatatgc ggtgtgaaat accgcacaga tgcgtaagga
     8221 gaaaataccg catcaggcgc cattcgccat tcaggctgcg caactgttgg gaagggcgat
     8281 cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat
     8341 taagttgggt aacgccaggg ttttcccagt cacgacgttg taaaacgacg gccagtgcca
     8401 agctg
//
