LOCUS       sequence-376965-        7749 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For expression coding sequencing of CLSTN2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7749)
  AUTHORS   Xu P, He H, Gao Q, Zhou Y, Wu Z, Zhang X, Sun L, Hu G, Guan Q, You 
            Z, Zhang X, Zheng W, Xiong M, Chen Y
  TITLE     Human midbrain dopaminergic neuronal differentiation markers predict
            cell therapy outcomes in a Parkinson's disease model.
  JOURNAL   J Clin Invest. 2022 Jul 15;132(14). pii: 156768. doi: 
            10.1172/JCI156768.
  PUBMED    35700056
REFERENCE   2  (bases 1 to 7749)
  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: "J Clin 
            Invest."; date: "2022-07-15"; volume: "132(14). pii: 156768. doi"; 
            pages: " 10.1172/JCI156768"
COMMENT     Sequence Label: pCAG-3xHAtag-SP-CLSTN2
FEATURES             Location/Qualifiers
     source          1..7749
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        5..384
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          662..1678
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1602..1624
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1686..1705
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             1737..1826
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAGYPYDVPDYAGSYPYDVPDYA"
     primer_bind     complement(4771..4790)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4836..4891
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4890..4909)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5252..5268)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5252..5268)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5265..5287)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5276..5292
                     /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(5300..5306,5307..5324,5325..5330))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5345..5366
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5425..5620
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      5471..5606
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    5626..5760
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5663..5682)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5717..5736
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5828..5845)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5999..6587)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6079..6098)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6758..7549,7550..7618))
                     /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     7381..7400
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7619..7723)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ggtcgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat
       61 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg
      121 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
      181 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
      241 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
      301 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
      361 gtattagtca tcgctattac catggtcgag gtgagcccca cgttctgctt cactctcccc
      421 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca
      481 gcgatggggg cggggggggg gggggsgcgc gccrggsggg gggggggggg gggggggggg
      541 gggsggggsg rggsggagrg gkgcggcggc agccaatcag agcggcgcgc tccgaaagtt
      601 tccttttatg gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc
      661 gggagtcgct gcgcgctgcc ttcgccccgt gccccgctcc gccgccgcct cgcgccgccc
      721 gccccggctc tgactgaccg cgttactccc acaggtgagc gggcgggacg gcccttctcc
      781 tccgggctgt aattagcgct tggtttaatg acggcttgtt tcttttctgt ggctgcgtga
      841 aagccttgag gggctccggg agggcccttt gtgcgggggg agcggctcgg ggggtgcgtg
      901 cgtgtgtgtg tgcgtgggga gcgccgcgtg cggctccgcg ctgcccggcg gctgtgagcg
      961 ctgcgggcgc ggcgcggggc tttgtgcgct ccgcagtgtg cgcgagggga gcgcggccgg
     1021 gggcggtgcc ccgcggtgcg gggggggctg cgaggggaac aaaggctgcg tgcggggtgt
     1081 gtgcgtgggg gggtgagcag ggggtgtggg cgcgtcggtc gggctgcaac cccccctgca
     1141 cccccctccc cgagttgctg agcacggccc ggcttcgggt gcggggctcc gtacggggcg
     1201 tggcgcgggg ctcgccgtgc cgggcggggg gtggcggcag gtgggggtgc cgggcggggc
     1261 ggggccgcct cgggccgggg rgggctcggg ggrggggcgc ggcggccccc ggagcgccgg
     1321 cggctgtcga ggcgcggcga gccgcagcca ttgcctttta tggtaatcgt gcgagagggc
     1381 gcagggactt cctttgtccc aaatctgtgc ggagccgaaa tctgggaggc gccgccgcac
     1441 cccctctagc gggcgcgggg cgaagcggtg cggcgccggc aggaaggaaa tgggcgggga
     1501 gggccttcgt gcgtcgccgc gccgccgtcc ccttctccct ctccagcctc ggggctgtcc
     1561 gcggggggac ggctgccttc gggggggacg gggcagggcg gggttcggct tctggcgtgt
     1621 gaccggcggc tctagagcct ctgctaacca tgttcatgcc ttcttctttt tcctacagct
     1681 cctgggcaac gtgctggtta ttgtgctgtc tcatcatttt ggcaaaggcc accatgtacc
     1741 catacgatgt tccagattac gctggctatc cctatgacgt cccggactat gcaggatcct
     1801 atccatatga cgttccagat tacgctctgc ctgggcggct gtgctgggtg ccgctcctgc
     1861 tggcgctggg cgtggggagc ggcagcggcg gtggcgggga cagccggcag cgccgcctcc
     1921 tcgcggctaa agtcaataag cacaagccat ggatcgagac ttcatatcat ggagtcataa
     1981 ctgagaacaa tgacacagtc attttggacc caccactggt agccctggat aaagatgcac
     2041 cggttccttt tgcaggggaa atctgtgcgt tcaagatcca tggccaggag ctgccctttg
     2101 aggctgtggt gctcaacaag acatcaggag agggccggct ccgtgccaag agccccattg
     2161 actgtgagtt gcagaaggag tacacattca tcatccaggc ctatgactgt ggtgctgggc
     2221 cccacgagac agcctggaaa aagtcacaca aggccgtggt ccatatacag gtgaaggatg
     2281 tcaacgagtt tgctcccacc ttcaaagagc cagcctacaa ggctgttgtg acggagggca
     2341 agatctatga cagcattctg caggtggagg ccattgacga ggactgctcc ccacagtaca
     2401 gccagatctg caactatgaa atcgtcacca cagatgtgcc ttttgccatc gacagaaatg
     2461 gcaacatcag gaacactgag aagctgagct atgacaaaca acaccagtat gagatcctgg
     2521 tgaccgccta cgactgtgga cagaagcccg ctgctcagga caccctggtg caggtggatg
     2581 tgaagccagt ttgcaagcct ggctggcaag actggaccaa gaggattgag taccagcctg
     2641 gctccgggag catgcccctg ttccccagca tccacctgga gacgtgcgat ggagccgtgt
     2701 cttccctcca gatcgtcaca gagctgcaga ctaattacat tgggaagggt tgtgaccggg
     2761 agacctactc tgagaaatcc cttcagaagt tatgtggagc ctcctctggc atcattgacc
     2821 tcttgccatc ccctagcgct gccaccaact ggactgcagg actgctggtg gacagcagtg
     2881 agatgatctt caagtttgac ggcaggcagg gtgccaaaat ccccgatggg attgtgccca
     2941 agaacctgac cgatcagttc accatcacca tgtggatgaa acacggcccc agccctggtg
     3001 tgagagccga gaaggaaacc atcctctgca actcagacaa aaccgaaatg aaccggcatc
     3061 actatgccct gtatgtgcac aactgccgcc tcgtctttct cttgcggaag gacttcgacc
     3121 aggctgacac ctttcgcccc gcggagttcc actggaagct ggatcagatt tgtgacaaag
     3181 agtggcacta ctatgtcatc aatgtggagt ttcctgtggt aaccttatac atggatggag
     3241 caacatatga accatacctg gtgaccaacg actggcccat tcatccatct cacatagcca
     3301 tgcaactcac agtcggcgct tgttggcaag gaggagaagt caccaaacca cagtttgctc
     3361 agttctttca tggaagcctg gccagtctca ccatccgccc tggcaaaatg gaaagccaga
     3421 aggtgatctc ctgcctgcag gcctgcaagg aagggctgga cattaattcc ttggaaagcc
     3481 ttggccaagg aataaagtat cacttcaacc cctcgcagtc catcctggtg atggaaggtg
     3541 acgacattgg gaacattaac cgtgctctcc agaaagtctc ctacatcaac tccaggcagt
     3601 tcccaacggc gggtgtgcgg cgcctcaaag tatcctccaa agtccagtgc tttggggaag
     3661 acgtatgcat cagtatccct gaggtagatg cctatgtgat ggtcctccag gccatcgagc
     3721 cccggatcac cctccggggc acagaccact tctggagacc tgctgcccag tttgaaagtg
     3781 ccaggggagt gaccctcttc cctgatatca agattgtgag caccttcgcc aaaaccgaag
     3841 cccccgggga cgtgaaaacc acagacccca aatcagaagt cttagaggaa atgcttcata
     3901 acttagattt ctgtgacatt ttggtgatcg gaggggactt ggacccaagg caggagtgct
     3961 tggagctcaa ccacagtgag ctccaccaac gacacctgga tgccactaat tctactgcag
     4021 gctactccat ctacggtgtg ggctccatga gccgctatga gcaggtgcta catcacatcc
     4081 gctaccgcaa ctggcgtccg gcttcccttg aggcccggcg tttccggatt aagtgctcag
     4141 aactcaatgg gcgctacact agcaatgagt tcaacttgga ggtcagcatc cttcatgaag
     4201 accaagtctc agataaggag catgtcaatc atctgattgt gcagcctccc ttcctccagt
     4261 ctgtccatca tcctgagtcc cggagtagca tccagcacag ttcagtggtc ccaagcattg
     4321 ccacagtggt catcatcatc tccgtgtgca tgcttgtgtt tgtcgtggcc atgggtgtgt
     4381 accgggtccg gatcgcccac cagcacttca tccaggagac tgaggctgcc aaggaatctg
     4441 agatggactg ggacgattct gcgctgacta tcacagtcaa ccccatggag aaacatgaag
     4501 gaccagggca tggggaagat gagactgagg gagaagagga ggaagaagcc gaggaagaaa
     4561 tgagctccag cagtggctct gacgacagcg aagaggagga ggaggaggaa gggatgggca
     4621 gaggcagaca tgggcagaat ggagccaggc aagcccagct ggagtgggat gactccaccc
     4681 tcccctacta ggcactcctc aggtgcaggc tgcctatcag aaggtggtgg ctggtgtggc
     4741 caatgccctg gctcacaaat accactgaga tctttttccc tctgccaaaa attatgggga
     4801 catcatgaag ccccttgagc atctgacttc tggctaataa aggaaattta ttttcattgc
     4861 aatagtgtgt tggaattttt tgtgtctctc actcggaagg acatatggga gggcaaatca
     4921 tttaaaacat cagaatgagt atttggttta gagtttggca acatatgccc atatgctggc
     4981 tgccatgaac aaaggttggc tataaagagg tcatcagtat atgaaacagc cccctgctgt
     5041 ccattcctta ttccatagaa aagccttgac ttgaggttag atttttttta tattttgttt
     5101 tgtgttattt ttttctttaa catccctaaa attttcctta catgttttac tagccagatt
     5161 tttcctcctc tcctgactac tcccagtcat agctgtccct cttctcttat ggagatccct
     5221 cgacctgcag cccaagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
     5281 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
     5341 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     5401 gaaacctgtc gtgccagcgg atccgcatct caattagtca gcaaccatag tcccgcccct
     5461 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg
     5521 actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa
     5581 gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctaactt gtttattgca
     5641 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     5701 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggatc
     5761 cgctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     5821 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     5881 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     5941 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     6001 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     6061 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     6121 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     6181 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     6241 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     6301 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     6361 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     6421 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     6481 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     6541 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     6601 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     6661 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     6721 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     6781 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     6841 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     6901 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     6961 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     7021 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     7081 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     7141 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     7201 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     7261 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     7321 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     7381 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     7441 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     7501 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     7561 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     7621 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     7681 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     7741 tgccacctg
//
