LOCUS       sequence-129653-        5627 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5627)
  AUTHORS   Prakriya M, Feske S, Gwack Y, Srikanth S, Rao A, Hogan PG
  TITLE     Orai1 is an essential pore subunit of the CRAC channel.
  JOURNAL   Nature. 2006 Sep 14. 443(7108):230-3.
  PUBMED    16921383
REFERENCE   2  (bases 1 to 5627)
  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: "Nature. 
            2006 Sep 14. 443(7108):230-3."
COMMENT     Sequence Label: Orai1 CFP
FEATURES             Location/Qualifiers
     source          1..5627
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        68..371
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        372..575
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     526..546
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      924..933
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      1574..1583
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1580..1582,1583..1585,1586..2299)
                     /codon_start=1
                     /product="enhanced CFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mECFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1625..1646)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1886..1905)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2233..2254
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2422..2543
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2459..2478)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2513..2532
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2550..3005)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2687..2708)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2899..2918
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3032..3136
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3138..3495
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3138..3329
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3161..3181)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3346..3481
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3530..4324
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             3530..4324
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3584..3603)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4194..4213
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4512..4531)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4556..4603
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4932..5520
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5421..5440
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 catgcattag ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg
       61 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
      121 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt
      181 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
      241 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
      301 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
      361 ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact
      421 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
      481 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta
      541 ggcgtgtacg gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatccgct
      601 agcgctaccg gactcagatc tcgagatgca tccggagccc gccccgcccc cgagccgtag
      661 cagtcccgag cttcccccaa gcggcggcag caccaccagc ggcagccgcc ggagccgccg
      721 ccgcagcggg gacggggagc ccccgggggc cccgccaccg ccgccgtccg ccgtcaccta
      781 cccggactgg atcggccaga gttactccga ggtgatgagc ctcaacgagc actccatgca
      841 ggcgctgtcc tggcgcaagc tctacttgag ccgcgccaag cttaaagcct ccagccggac
      901 ctcggctctg ctctccggct tcgccatggt ggcaatggtg gaggtgcagc tggacgctga
      961 ccacgactac ccaccggggc tgctcatcgc cttcagtgcc tgcaccacag tgctggtggc
     1021 tgtgcacctg tttgcgctca tgatcagcac ctgcatcctg cccaacatcg aggcggtgag
     1081 caacgtgcac aatctcaact cggtcaagga gtccccccat gagcgcatgc accgccacat
     1141 cgagctggcc tgggccttct ccaccgtcat tggcacgctg ctcttcctag ctgaggtggt
     1201 gctgctctgc tgggtcaagt tcttgcccct caagaagcag ccaggccagc caaggcccac
     1261 cagcaagccc cccgccagtg gcgcagcagc caacgtcagc accagcggca tcaccccggg
     1321 ccaggcagct gccatcgcct cgaccaccat catggtgccc ttcggcctga tctttatcgt
     1381 cttcgccgtc cacttctacc gctcactggt tagccataag accgaccgac agttccagga
     1441 gctcaacgag ctggcggagt ttgcccgctt acaggaccag ctggaccaca gaggggacca
     1501 ccccctgacg cccggcagcc actatgcccg aattctgcag tcgacggtac cgcgggcccg
     1561 ggatccaccg gtcgccacca tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc
     1621 catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg
     1681 cgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct
     1741 gcccgtgccc tggcccaccc tcgtgaccac cctgacctgg ggcgtgcagt gcttcagccg
     1801 ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt
     1861 ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa
     1921 gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga
     1981 cggcaacatc ctggggcaca agctggagta caactacatc agccacaacg tctatatcac
     2041 cgccgacaag cagaagaacg gcatcaaggc caacttcaag atccgccaca acatcgagga
     2101 cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt
     2161 gctgctgccc gacaaccact acctgagcac ccagtccaag ctgagcaaag accccaacga
     2221 gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat
     2281 ggacgagctg tacaagtaaa gcggccgcga ctctagatca taatcagcca taccacattt
     2341 gtagaggttt tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa
     2401 atgaatgcaa ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc
     2461 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     2521 tccaaactca tcaatgtatc ttaaggcgta aattgtaagc gttaatattt tgttaaaatt
     2581 cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat
     2641 cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa
     2701 gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg
     2761 cgatggccca ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa
     2821 agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc
     2881 gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag
     2941 tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg
     3001 cgcgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     3061 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     3121 ttgaaaaagg aagagtcctg aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg
     3181 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     3241 tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     3301 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact
     3361 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
     3421 gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
     3481 ctaggctttt gcaaagatcg atcaagagac aggatgagga tcgtttcgca tgattgaaca
     3541 agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg
     3601 ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg
     3661 cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aagacgaggc
     3721 agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt
     3781 cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc
     3841 atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca
     3901 tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc
     3961 acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg
     4021 gctcgcgcca gccgaactgt tcgccaggct caaggcgagc atgcccgacg gcgaggatct
     4081 cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc
     4141 tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc
     4201 tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta
     4261 cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt
     4321 ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga
     4381 gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac
     4441 gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccctagg
     4501 gggaggctaa ctgaaacacg gaaggagaca ataccggaag gaacccgcgc tatgacggca
     4561 ataaaaagac agaataaaac gcacggtgtt gggtcgtttg ttcataaacg cggggttcgg
     4621 tcccagggct ggcactctgt cgatacccca ccgagacccc attggggcca atacgcccgc
     4681 gtttcttcct tttccccacc ccacccccca agttcgggtg aaggcccagg gctcgcagcc
     4741 aacgtcgggg cggcaggccc tgccatagcc tcaggttact catatatact ttagattgat
     4801 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
     4861 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     4921 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     4981 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     5041 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
     5101 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     5161 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     5221 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     5281 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     5341 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     5401 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     5461 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     5521 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     5581 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgc
//
