LOCUS       sequence-376912-        8532 bp    DNA     circular SYN 14-APR-2025
DEFINITION  For expression coding sequencing of PTPRO.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8532)
  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 8532)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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-PTPRO
FEATURES             Location/Qualifiers
     source          1..8532
                     /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(5554..5573)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    5619..5674
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(5673..5692)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(6035..6051)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6035..6051)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6048..6070)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6059..6075
                     /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(6083..6089,6090..6107,6108..6113))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6128..6149
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        6208..6403
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      6254..6389
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    6409..6543
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6446..6465)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6500..6519
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6611..6628)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6782..7370)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6862..6881)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7541..8332,8333..8401))
                     /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     8164..8183
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8402..8506)
                     /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 gggggggcgc gccrgggggg gggggggggg gggggggggg
      541 gggsggggsg rggsggrgrg 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 tacgctgggc acctgcccac ggggatacac ggcgcccgcc
     1861 gcctcctgcc tctgctctgg ctctttgtgc tgttcaagaa tgctacagct ttccatgtaa
     1921 ctgtccaaga tgataataac atcgttgtct cattagaagc ttcagacgtc atcagtccag
     1981 catctgtgta tgttgtgaag ataactggtg aatccaaaaa ttatttcttc gaatttgagg
     2041 aattcaacag cactttgcct cctcctgtta ttttcaaggc cagttatcat ggcctttatt
     2101 atataatcac tctggtagtg gtaaatggaa atgtggtgac caagccatcc agatcaatca
     2161 ctgtgttaac aaaacctcta cctgtaacca gtgtttccat atatgactat aaaccttctc
     2221 ctgaaacagg agtcctgttt gaaatacatt atccagaaaa atataacgtt ttcacaagag
     2281 tgaacattag ctactgggaa ggtaaagact tccggacaat gctatataaa gatttcttta
     2341 agggaaaaac agtatttaat cactggctgc caggaatgtg ttatagtaat atcacctttc
     2401 agctggtatc tgaggcaact tttaataaaa gtacccttgt tgagtacagt ggtgtcagtc
     2461 acgaacccaa acagcacaga actgcccctt atccacctca aaatatttcc gttcgtatcg
     2521 taaacttgaa caaaaacaac tgggaagaac agagtggcaa tttcccagaa gaatccttca
     2581 tgagatcaca agatacaata ggaaaagaaa aactcttcca ttttacagaa gaaacccctg
     2641 aaattccctc gggcaacatt tcttccggtt ggcctgattt taatagcagt gactatgaaa
     2701 ctacgtctca gccatattgg tgggacagtg catctgcagc tcctgaaagt gaagatgaat
     2761 ttgtcagcgt acttcccatg gaatacgaaa ataacagtac actcagtgag acagagaagt
     2821 caacatcagg ctctttctcc tttttccctg tgcaaatgat attgacctgg ttaccaccca
     2881 aaccacccac tgcttttgat gggttccata tccatattga acgagaagag aactttactg
     2941 aatatttgat ggtggatgaa gaagcacatg aatttgttgc agaactgaag gaacctggga
     3001 aatataagtt atctgtgaca acctttagtt cctcaggatc ttgtgaaact cgaaaaagtc
     3061 agtcagcaaa atcactcagc ttttatatca gtccttcagg agagtggatt gaagaactga
     3121 ccgagaagcc gcagcacgtg agtgtccacg ttttaagctc aaccactgcc ttgatgtcct
     3181 ggacatcttc ccaagagaac tacaacagca ccattgtgtc tgtggtgtcg ctgacctgcc
     3241 agaaacaaaa ggagagccag aggcttgaaa agcagtactg cactcaggtg aactcaagca
     3301 aacctattat tgaaaatctg gttcctggtg cccagtacca ggttgtaata tacctaagga
     3361 aaggcccttt gattggacca ccttcagatc ctgtgacatt tgctattgtt cccacaggaa
     3421 taaaggattt aatgctctat cctttgggtc ctacggccgt ggttctgagc tggaccagac
     3481 cttatttagg cgtgttcaga aaatacgtgg ttgaaatgtt ttatttcaac cctgctacaa
     3541 tgacatcaga gtggaccacc tactatgaaa tagcagcaac tgtttcctta actgcatccg
     3601 tgagaatagc taatctgctg ccagcatggt actacaactt ccgggttacc atggtgacgt
     3661 ggggagatcc agaattgagc tgctgtgaca gctctaccat cagcttcata acagccccag
     3721 tggctccgga aatcacttct gtggaatatt tcaacagtct gttatatatc agttggacat
     3781 atggggatga tacaacggac ttgtcccatt ctagaatgct tcactggatg gtggttgcag
     3841 aaggaaaaaa gaaaattaaa aagagtgtaa cacgcaatgt catgactgca attctcagct
     3901 tgcctccagg cgacatctat aacctctcag taactgcttg tactgaaaga ggaagtaata
     3961 cctccatgct ccgccttgtc aagctagaac cagctccacc caaatcactc ttcgcagtga
     4021 acaaaaccca gacttcagtg actttgctgt gggtggaaga gggagtagct gatttctttg
     4081 aagttttctg tcaacaagtt ggctccagtc agaaaaccaa acttcaggaa ccagttgctg
     4141 tttcttccca tgtcgtgacc atctccagcc ttcttcctgc cactgcctac aattgtagtg
     4201 tcaccagctt tagccatgac agccccagtg tccctacgtt catagccgtc tcaacaatgg
     4261 ttacagagat gaatcccaat gtggtagtga tctccgtgct ggccatcctt agcacacttt
     4321 taattggact gttgcttgtt accctcatta ttcttaggaa aaagcatctg cagatggcta
     4381 gggagtgtgg agctggtaca tttgtcaatt ttgcatcctt agagagggat ggaaagcttc
     4441 catacaactg gcgtaggagt atatttgctt tcttaaccct gctaccctca tgtctttgga
     4501 ctgattatct tttggcattt tatattaatc cttggagtaa aaatggttta aagaagagga
     4561 aactgacaaa cccggttcaa ctggatgact ttgatgccta tattaaggat atggccaaag
     4621 actctgacta taaattttct cttcagtttg aggagttgaa attgattgga ctggatatcc
     4681 cacactttgc tgcagatctt ccactgaatc gatgtaaaaa ccgttacaca aacatcctac
     4741 catatgactt cagccgtgtg agattagtct ccatgaatga agaggaaggt gcagactaca
     4801 tcaatgccaa ctatattcct ggatacaact caccccagga gtatattgcc acccaggggc
     4861 cactgcctga aaccagaaat gacttctgga agatggtcct gcaacaaaag tctcagatta
     4921 ttgtcatgct cactcagtgt aatgagaaaa ggagggtgaa atgtgaccat tactggccat
     4981 tcacggaaga acctatagcc tatggagaca tcactgtgga gatgatttca gaggaagagc
     5041 aggacgactg ggcctgtaga cacttccgga tcaactatgc tgacgagatg caggatgtga
     5101 tgcattttaa ctacactgca tggcctgatc atggtgtgcc cacagcaaat gctgcagaaa
     5161 gtatcctgca gtttgtacac atggtccgac agcaagctac caagagcaaa ggtcccatga
     5221 tcattcactg cagtgctggc gtgggacgga caggaacatt cattgccctg gacaggctct
     5281 tgcagcacat tcgggatcat gagtttgttg acatcttagg gctggtgtca gaaatgaggt
     5341 cataccggat gtctatggta cagacagagg agcagtacat ttttatccat cagtgtgtgc
     5401 aactgatgtg gatgaagaag aagcagcagt tctgcatcag tgatgtcata tacgagaatg
     5461 ttagcaagtc ctaggcactc ctcaggtgca ggctgcctat cagaaggtgg tggctggtgt
     5521 ggccaatgcc ctggctcaca aataccactg agatcttttt ccctctgcca aaaattatgg
     5581 ggacatcatg aagccccttg agcatctgac ttctggctaa taaaggaaat ttattttcat
     5641 tgcaatagtg tgttggaatt ttttgtgtct ctcactcgga aggacatatg ggagggcaaa
     5701 tcatttaaaa catcagaatg agtatttggt ttagagtttg gcaacatatg cccatatgct
     5761 ggctgccatg aacaaaggtt ggctataaag aggtcatcag tatatgaaac agccccctgc
     5821 tgtccattcc ttattccata gaaaagcctt gacttgaggt tagatttttt ttatattttg
     5881 ttttgtgtta tttttttctt taacatccct aaaattttcc ttacatgttt tactagccag
     5941 atttttcctc ctctcctgac tactcccagt catagctgtc cctcttctct tatggagatc
     6001 cctcgacctg cagcccaagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt
     6061 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg
     6121 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt
     6181 cgggaaacct gtcgtgccag cggatccgca tctcaattag tcagcaacca tagtcccgcc
     6241 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
     6301 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca
     6361 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctaa cttgtttatt
     6421 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     6481 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg
     6541 atccgctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     6601 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     6661 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     6721 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     6781 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     6841 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     6901 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     6961 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     7021 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     7081 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     7141 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     7201 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     7261 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     7321 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     7381 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     7441 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     7501 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     7561 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     7621 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     7681 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     7741 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     7801 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     7861 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     7921 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     7981 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     8041 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     8101 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     8161 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     8221 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     8281 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     8341 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     8401 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     8461 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     8521 aagtgccacc tg
//
