LOCUS       sequence-80941-e        8222 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human GPR56 and GFP in mammalian cells..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8222)
  AUTHORS   Bae BI, Tietjen I, Atabay KD, Evrony GD, Johnson MB, Asare E, Wang 
            PP, Murayama AY, Im K, Lisgo SN, Overman L, Sestan N, Chang BS, 
            Barkovich AJ, Grant PE, Topcu M, Politsky J, Okano H, Piao X, Walsh 
            CA
  TITLE     Evolutionarily dynamic alternative splicing of GPR56 regulates 
            regional cerebral cortical patterning.
  JOURNAL   Science. 2014 Feb 14;343(6172):764-8. doi: 10.1126/science.1244392.
  PUBMED    24531968
REFERENCE   2  (bases 1 to 8222)
  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: "10.1126/science"; 
            journalName: "Science"; date: "2014-02-14- 14"; volume: "343"; 
            issue: "6172"; pages: "764-8"
COMMENT     Sequence Label: pCAG-hGPR56-IRES-GFP
FEATURES             Location/Qualifiers
     source          1..8222
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        4..383
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        386..661
                     /label=chicken beta-actin promoter
     intron          662..1670
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1594..1616
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     regulatory      3507..3516
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    join(3854..4428,4429..4431,4432..4440)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4021..4038)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     4248..4267
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(4441..4443,4444..4446,4447..5160)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(4441..4443,4444..4446,4447..5157)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4486..4507)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4747..4766)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5094..5115
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(5246..5265)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    5311..5366
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(5365..5384)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5725..5741)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5725..5741)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5738..5760)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5749..5765
                     /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(5773..5779,5780..5797,5798..5803))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5818..5839
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5898..6093
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      5944..6079
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    6099..6233
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6136..6155)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6190..6209
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6301..6318)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6472..7060)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6552..6571)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7231..8022,8023..8091))
                     /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     7854..7873
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8092..8196)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
       61 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
      121 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
      181 ggactttcca ttgacgtcaa tgggtggact atttacggta aactgcccac ttggcagtac
      241 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
      301 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
      361 tattagtcat cgctattacc atgggtcgag gtgagcccca cgttctgctt cactctcccc
      421 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca
      481 gcgatggggg cggggggggg gggggcgcgc gccaggcggg gcggggcggg gcgaggggcg
      541 gggcggggcg aggcggagag gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt
      601 tccttttatg gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc
      661 gggagtcgct gcgttgcctt cgccccgtgc cccgctccgc gccgcctcgc gccgcccgcc
      721 ccggctctga ctgaccgcgt tactcccaca ggtgagcggg cgggacggcc cttctcctcc
      781 gggctgtaat tagcgcttgg tttaatgacg gctcgtttct tttctgtggc tgcgtgaaag
      841 ccttaaaggg ctccgggagg gccctttgtg cgggggggag cggctcgggg ggtgcgtgcg
      901 tgtgtgtgtg cgtggggagc gccgcgtgcg gcccgcgctg cccggcggct gtgagcgctg
      961 cgggcgcggc gcggggcttt gtgcgctccg cgtgtgcgcg aggggagcgc ggccgggggc
     1021 ggtgccccgc ggtgcggggg ggctgcgagg ggaacaaagg ctgcgtgcgg ggtgtgtgcg
     1081 tgggggggtg agcagggggt gtgggcgcgg cggtcgggct gtaacccccc cctgcacccc
     1141 cctccccgag ttgctgagca cggcccggct tcgggtgcgg ggctccgtgc ggggcgtggc
     1201 gcggggctcg ccgtgccggg cggggggtgg cggcaggtgg gggtgccggg cggggcgggg
     1261 ccgcctcggg ccggggaggg ctcgggggag gggcgcggcg gccccggagc gccggcggct
     1321 gtcgaggcgc ggcgagccgc agccattgcc ttttatggta atcgtgcgag agggcgcagg
     1381 gacttccttt gtcccaaatc tggcggagcc gaaatctggg aggcgccgcc gcaccccctc
     1441 tagcgggcgc gggcgaagcg gtgcggcgcc ggcaggaagg aaatgggcgg ggagggcctt
     1501 cgtgcgtcgc cgcgccgccg tccccttctc catctccagc ctcggggctg ccgcaggggg
     1561 acggctgcct tcggggggga cggggcaggg cggggttcgg cttctggcgt gtgaccggcg
     1621 gctctagagc ctctgctaac catgttcatg ccttcttctt tttcctacag ctcctgggca
     1681 acgtgctggt tgttgtgctg tctcatcatt ttggcaaaga attcctcgag gatgccacca
     1741 tgactcccca gtcgctgctg cagacgacac tgttcctgct gagtctgctc ttcctggtcc
     1801 aaggtgccca cggcaggggc cacagggaag actttcgctt ctgcagccag cggaaccaga
     1861 cacacaggag cagcctccac tacaaaccca caccagacct gcgcatctcc atcgagaact
     1921 ccgaagaggc cctcacagtc catgcccctt tccctgcagc ccaccctgct tcccgatcct
     1981 tccctgaccc caggggcctc taccacttct gcctctactg gaaccgacat gctgggagat
     2041 tacatcttct ctatggcaag cgtgacttct tgctgagtga caaagcctct agcctcctct
     2101 gcttccagca ccaggaggag agcctggctc agggcccccc gctgttagcc acttctgtca
     2161 cctcctggtg gagccctcag aacatcagcc tgcccagtgc cgccagcttc accttctcct
     2221 tccacagtcc tccccacacg gccgctcaca atgcctcggt ggacatgtgc gagctcaaaa
     2281 gggacctcca gctgctcagc cagttcctga agcatcccca gaaggcctca aggaggccct
     2341 cggctgcccc cgccagccag cagttgcaga gcctggagtc gaaactgacc tctgtgagat
     2401 tcatggggga catggtgtcc ttcgaggagg accggatcaa cgccacggtg tggaagctcc
     2461 agcccacagc cggcctccag gacctgcaca tccactcccg gcaggaggag gagcagagcg
     2521 agatcatgga gtactcggtg ctgctgcctc gaacactctt ccagaggacg aaaggccgga
     2581 gcggggaggc tgagaagaga ctcctcctgg tggacttcag cagccaagcc ctgttccagg
     2641 acaagaattc cagccaagtc ctgggtgaga aggtcttggg gattgtggta cagaacacca
     2701 aagtagccaa cctcacggag cccgtggtgc tcaccttcca gcaccagcta cagccgaaga
     2761 atgtgactct gcaatgtgtg ttctgggttg aagaccccac attgagcagc ccggggcatt
     2821 ggagcagtgc tgggtgtgag accgtcagga gagaaaccca aacatcctgc ttctgcaacc
     2881 acttgaccta ctttgcagtg ctgatggtct cctcggtgga ggtggacgcc gtgcacaagc
     2941 actacctgag cctcctctcc tacgtgggct gtgtcgtctc tgccctggcc tgccttgtca
     3001 ccattgccgc ctacctctgc tccagggtgc ccctgccgtg caggaggaaa cctcgggact
     3061 acaccatcaa ggtgcacatg aacctgctgc tggccgtctt cctgctggac acgagcttcc
     3121 tgctcagcga gccggtggcc ctgacaggct ctgaggctgg ctgccgagcc agtgccatct
     3181 tcctgcactt ctccctgctc acctgccttt cctggatggg cctcgagggg tacaacctct
     3241 accgactcgt ggtggaggtc tttggcacct atgtccctgg ctacctactc aagctgagcg
     3301 ccatgggctg gggcttcccc atctttctgg tgacgctggt ggccctggtg gatgtggaca
     3361 actatggccc catcatcttg gctgtgcata ggactccaga gggcgtcatc tacccttcca
     3421 tgtgctggat ccgggactcc ctggtcagct acatcaccaa cctgggcctc ttcagcctgg
     3481 tgtttctgtt caacatggcc atgctagcca ccatggtggt gcagatcctg cggctgcgcc
     3541 cccacaccca aaagtggtca catgtgctga cactgctggg cctcagcctg gtccttggcc
     3601 tgccctgggc cttgatcttc ttctcctttg cttctggcac cttccagctt gtcgtcctct
     3661 accttttcag catcatcacc tccttccaag gcttcctcat cttcatctgg tactggtcca
     3721 tgcggctgca ggcccggggt ggcccctccc ctctgaagag caactcagac agcgccaggc
     3781 tccccatcag ctcgggcagc acctcgtcca gccgcatcta atcgcggccg ctacgtaaat
     3841 tccgcccccc ccccccctct ccctcccccc cccctaacgt tactggccga agccgcttgg
     3901 aataaggccg gtgtgcgttt gtctatatgt tattttccac catattgccg tcttttggca
     3961 atgtgagggc ccggaaacct ggccctgtct tcttgacgag cattcctagg ggtctttccc
     4021 ctctcgccaa aggaatgcaa ggtctgttga atgtcgtgaa ggaagcagtt cctctggaag
     4081 cttcttgaag acaaacaacg tctgtagcga ccctttgcag gcagcggaac cccccacctg
     4141 gcgacaggtg cctctgcggc caaaagccac gtgtataaga tacacctgca aaggcggcac
     4201 aaccccagtg ccacgttgtg agttggatag ttgtggaaag agtcaaatgg ctctcctcaa
     4261 gcgtattcaa caaggggctg aaggatgccc agaaggtacc ccattgtatg ggatctgatc
     4321 tggggcctcg gtgcacatgc tttacatgtg tttagtcgag gttaaaaaaa cgtctaggcc
     4381 ccccgaacca cggggacgtg gttttccttt gaaaaacacg atgataatat ggccacaacc
     4441 atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac
     4501 ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac
     4561 ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc
     4621 ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc gctaccccga ccacatgaag
     4681 cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc
     4741 ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg
     4801 gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac
     4861 aagctggagt acaactacaa cagccacaac gtctatatca tggccgacaa gcagaagaac
     4921 ggcatcaagg tgaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc
     4981 gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac
     5041 tacctgagca cccagtccgc cctgagcaaa gaccccaacg agaagcgcga tcacatggtc
     5101 ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagtaa
     5161 agcggccggc cgccagcaca gtggtcgatc gacctgcagg catgcaagct tcagccaatg
     5221 ccctggctca caaataccac tgagatcttt ttccctctgc caaaaattat ggggacatca
     5281 tgaagcccct tgagcatctg acttctggct aataaaggaa atttattttc attgcaatag
     5341 tgtgttggaa ttttttgtgt ctctcactcg gaaggacata tgggagggca aatcatttaa
     5401 aacatcagaa tgagtatttg gtttagagtt tggcaacata tgccatatgc tggctgccat
     5461 gaacaaaggt ggctataaag aggtcatcag tatatgaaac agccccctgc tgtccattcc
     5521 ttattccata gaaaagcctt gacttgaggt tagatttttt ttatattttg ttttgtgtta
     5581 tttttttctt taacatccct aaaattttcc ttacatgttt tactagccag atttttcctc
     5641 ctctcctgac tactcccagt catagctgtc cctcttctct tatgaagatc cctcgacctg
     5701 cagcccaagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     5761 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     5821 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     5881 gtcgtgccag cggatccgca tctcaattag tcagcaacca tagtcccgcc cctaactccg
     5941 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
     6001 ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga
     6061 ggaggctttt ttggaggcct aggcttttgc aaaaagctaa cttgtttatt gcagcttata
     6121 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     6181 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg atccgctgca
     6241 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     6301 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     6361 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     6421 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     6481 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     6541 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     6601 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     6661 ttctcaatgc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     6721 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     6781 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     6841 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     6901 ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     6961 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     7021 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     7081 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     7141 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     7201 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     7261 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     7321 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     7381 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     7441 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     7501 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     7561 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     7621 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     7681 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     7741 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     7801 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     7861 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     7921 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     7981 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     8041 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     8101 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     8161 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     8221 tg
//
