LOCUS       sequence-376968-        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-SP-CLSTN2-3xHAtag
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             4599..4688
                     /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 gccrgggggg gggggggggg gggggggggg
      541 gggsggggsg rggsggrgag 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 accatgctgc
     1741 ctgggcggct gtgctgggtg ccgctcctgc tggcgctggg cgtggggagc ggcagcggcg
     1801 gtggcgggga cagccggcag cgccgcctcc tcgcggctaa agtcaataag cacaagccat
     1861 ggatcgagac ttcatatcat ggagtcataa ctgagaacaa tgacacagtc attttggacc
     1921 caccactggt agccctggat aaagatgcac cggttccttt tgcaggggaa atctgtgcgt
     1981 tcaagatcca tggccaggag ctgccctttg aggctgtggt gctcaacaag acatcaggag
     2041 agggccggct ccgtgccaag agccccattg actgtgagtt gcagaaggag tacacattca
     2101 tcatccaggc ctatgactgt ggtgctgggc cccacgagac agcctggaaa aagtcacaca
     2161 aggccgtggt ccatatacag gtgaaggatg tcaacgagtt tgctcccacc ttcaaagagc
     2221 cagcctacaa ggctgttgtg acggagggca agatctatga cagcattctg caggtggagg
     2281 ccattgacga ggactgctcc ccacagtaca gccagatctg caactatgaa atcgtcacca
     2341 cagatgtgcc ttttgccatc gacagaaatg gcaacatcag gaacactgag aagctgagct
     2401 atgacaaaca acaccagtat gagatcctgg tgaccgccta cgactgtgga cagaagcccg
     2461 ctgctcagga caccctggtg caggtggatg tgaagccagt ttgcaagcct ggctggcaag
     2521 actggaccaa gaggattgag taccagcctg gctccgggag catgcccctg ttccccagca
     2581 tccacctgga gacgtgcgat ggagccgtgt cttccctcca gatcgtcaca gagctgcaga
     2641 ctaattacat tgggaagggt tgtgaccggg agacctactc tgagaaatcc cttcagaagt
     2701 tatgtggagc ctcctctggc atcattgacc tcttgccatc ccctagcgct gccaccaact
     2761 ggactgcagg actgctggtg gacagcagtg agatgatctt caagtttgac ggcaggcagg
     2821 gtgccaaaat ccccgatggg attgtgccca agaacctgac cgatcagttc accatcacca
     2881 tgtggatgaa acacggcccc agccctggtg tgagagccga gaaggaaacc atcctctgca
     2941 actcagacaa aaccgaaatg aaccggcatc actatgccct gtatgtgcac aactgccgcc
     3001 tcgtctttct cttgcggaag gacttcgacc aggctgacac ctttcgcccc gcggagttcc
     3061 actggaagct ggatcagatt tgtgacaaag agtggcacta ctatgtcatc aatgtggagt
     3121 ttcctgtggt aaccttatac atggatggag caacatatga accatacctg gtgaccaacg
     3181 actggcccat tcatccatct cacatagcca tgcaactcac agtcggcgct tgttggcaag
     3241 gaggagaagt caccaaacca cagtttgctc agttctttca tggaagcctg gccagtctca
     3301 ccatccgccc tggcaaaatg gaaagccaga aggtgatctc ctgcctgcag gcctgcaagg
     3361 aagggctgga cattaattcc ttggaaagcc ttggccaagg aataaagtat cacttcaacc
     3421 cctcgcagtc catcctggtg atggaaggtg acgacattgg gaacattaac cgtgctctcc
     3481 agaaagtctc ctacatcaac tccaggcagt tcccaacggc gggtgtgcgg cgcctcaaag
     3541 tatcctccaa agtccagtgc tttggggaag acgtatgcat cagtatccct gaggtagatg
     3601 cctatgtgat ggtcctccag gccatcgagc cccggatcac cctccggggc acagaccact
     3661 tctggagacc tgctgcccag tttgaaagtg ccaggggagt gaccctcttc cctgatatca
     3721 agattgtgag caccttcgcc aaaaccgaag cccccgggga cgtgaaaacc acagacccca
     3781 aatcagaagt cttagaggaa atgcttcata acttagattt ctgtgacatt ttggtgatcg
     3841 gaggggactt ggacccaagg caggagtgct tggagctcaa ccacagtgag ctccaccaac
     3901 gacacctgga tgccactaat tctactgcag gctactccat ctacggtgtg ggctccatga
     3961 gccgctatga gcaggtgcta catcacatcc gctaccgcaa ctggcgtccg gcttcccttg
     4021 aggcccggcg tttccggatt aagtgctcag aactcaatgg gcgctacact agcaatgagt
     4081 tcaacttgga ggtcagcatc cttcatgaag accaagtctc agataaggag catgtcaatc
     4141 atctgattgt gcagcctccc ttcctccagt ctgtccatca tcctgagtcc cggagtagca
     4201 tccagcacag ttcagtggtc ccaagcattg ccacagtggt catcatcatc tccgtgtgca
     4261 tgcttgtgtt tgtcgtggcc atgggtgtgt accgggtccg gatcgcccac cagcacttca
     4321 tccaggagac tgaggctgcc aaggaatctg agatggactg ggacgattct gcgctgacta
     4381 tcacagtcaa ccccatggag aaacatgaag gaccagggca tggggaagat gagactgagg
     4441 gagaagagga ggaagaagcc gaggaagaaa tgagctccag cagtggctct gacgacagcg
     4501 aagaggagga ggaggaggaa gggatgggca gaggcagaca tgggcagaat ggagccaggc
     4561 aagcccagct ggagtgggat gactccaccc tcccctacta cccatacgat gttccagatt
     4621 acgctggcta tccctatgac gtcccggact atgcaggatc ctatccatat gacgttccag
     4681 attacgctta 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
//
