LOCUS       216695       6212 bp DNA     circular SYN 20-APR-2023
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6212)
  AUTHORS   Octave
  TITLE     Direct Submission
  JOURNAL   Exported Feb 14, 2024 from SnapGene 7.1.1
            https://www.snapgene.com
COMMENT     Ligation of : Str-KDEL_ManII-SBP-mApple-FIREtag.xdna [5652 nt] : 
            (NotI[1578] / NheI[927]) to MATE-KDEL_SBP-mApple-FIREt ag-GPI.xdna 
            [552  nt] : (NheI[927] / NotI[1482])
FEATURES             Location/Qualifiers
     source          1..6212
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     863..882
                     /label=T7
                     /note="note"
                     /note="color: #008b8b; direction: RIGHT"
     misc_feature    954..1067
                     /label=STIM1 SP
                     /note="note"
                     /note="color: #996633; direction: RIGHT"
     misc_feature    954..1067
                     /label=STIM1 Signal peptide
                     /note="note"
                     /note="color: #996633; direction: RIGHT"
     misc_feature    1077..1415
                     /label=FIREmate
                     /note="color: #bd3d48; direction: RIGHT"
     CDS             1431..1457
                     /label=HA tag
                     /note="color: #008000; direction: RIGHT"
     misc_feature    1467..1478
                     /label=KDEL
                     /note="note=""direct strand 
                     translation""translation=""KDEL"""
                     /note="color: #ff00ff; direction: RIGHT"
     misc_feature    1467..1478
                     /label=KDEL
                     /note="color: #c0066e; direction: RIGHT"
     intron          1514..1809
                     /label=synth int
                     /note="note"
                     /note="color: #4682b4; direction: RIGHT"
     primer_bind     complement(1892..1910)
                     /label=\'a0pIRES-RP
                     /note="color: #ffff00; direction: LEFT"
     misc_feature    2437..2784
                     /label=insert from SBP-mApple.xdna
                     /label=insert from FseI-FIREtag-SfiI
                     /label=insert from MATE-KDEL_SBP-mApple-FIREtag-GPI.xdna
                     /note="note=""direct strand 
                     translation""translation=""MKLSRQFTVFGSAIFCVVIFSLYLMLDRGHLD
                     YPRGPRQEGSFPQGQLSILQEKIDHLERLLAENNEIISNIRDSVINLSESVEDGPRGSP
                     GNASQGSIHLHSPQLALQADPRDCL""note=""Fragment extracted from 
                     SBP-mApple.xdna [829  nt] : (EcoRI[1] / FseI[833]) 
                     3643..3695"
                     /note="note=""Fragment extracted from FseI-FIREtag-SfiI [53
                      nt] : (Uncut 5'[0] / Uncut 3'[53]) 932..1483"
                     /note="note=""Fragment extracted from 
                     MATE-KDEL_SBP-mApple-FIREtag-GPI.xdna [552  nt] : 
                     (NheI[927] / NotI[1482])"
                     /note="color: #84a4c0; direction: RIGHT"
     CDS             2812..2922
                     /label=SBP tag
                     /note="color: #008000; direction: RIGHT"
     misc_feature    2932..3636
                     /label=mApple
                     /note="note=""direct strand 
                     translation""translation=""VSKGEENNMAIIKEFMRFKVHMEGSVNGHEFE
                     IEGEGEGRPYEAFQTAKLKVTKGGPLPFAWDILSPQFMYGSKVYIKHPADIPDYFKLSF
                     PEGFRWERVMNFEDGGIIHVNQDSSLQDGVFIYKVKLRGTNFPSDGPVMQKKTMGWEAS
                     EERMYPEDGALKSEIKKRLKLKDGGHYAAEVKTTYKAKKPVQLPGAYIVDIKLDIVSHN
                     "
                     /note="color: #c03506; direction: RIGHT"
     misc_feature    3646..3660
                     /label=linker
                     /label=insert from PmeI-FIREmate-HA-RsrII.xdna
                     /note="note=""direct strand 
                     translation""translation=""GGGSG"" 1073..1461"
                     /note="note=""Fragment extracted from 
                     PmeI-FIREmate-HA-RsrII.xdna [389  nt] : (PmeI[2] / 
                     RsrII[393])"
                     /note="color: #2427bf; direction: RIGHT"
     misc_feature    3661..3691
                     /label=FIREtag[mod]
                     /note="color: #bd2c2d; direction: RIGHT"
     primer_bind     complement(3754..3773)
                     /label=\'a0pEGFP_C2-RP
                     /note="color: #ffff00; direction: LEFT"
     primer_bind     3923..3942
                     /label=EBV-RP
                     /note="color: #ffff00; direction: RIGHT"
     primer_bind     complement(4045..4065)
                     /label=RPC
                     /note="color: #ffff00; direction: LEFT"
     primer_bind     complement(4051..4071)
                     /label=M13-rev
                     /note="note"
                     /note="color: #008b8b; direction: LEFT"
     primer_bind     complement(4054..4071)
                     /label=M13-RP
                     /note="color: #ffff00; direction: LEFT"
     primer_bind     complement(4069..4088)
                     /label=\'a0pQE-FP
                     /note="color: #ffff00; direction: LEFT"
     misc_binding    complement(4077..4099)
                     /label=LacO
                     /note="note"
                     /note="color: #6495ed; direction: LEFT"
     primer_bind     complement(4093..4114)
                     /label=BAC1
                     /note="color: #ffff00; direction: LEFT"
     promoter        complement(4104..4133)
                     /label=lac
                     /note="note"
                     /note="color: #4682b4; direction: LEFT"
     primer_bind     complement(4144..4160)
                     /label=PCR2
                     /note="color: #ffff00; direction: LEFT"
     rep_origin      complement(4439..5067)
                     /direction=LEFT
                     /label=ColE1 origin
                     /note="note"
                     /note="color: #4682b4"
     CDS             complement(5219..5878)
                     /label=AmpR
                     /note="note"
                     /note="color: #8fbc8f; direction: LEFT"
     promoter        complement(6118..6146)
                     /label=Amp prom
                     /note="note"
                     /note="color: #4682b4; direction: LEFT"
     primer_bind     6192..6209
                     /label=pBR1
                     /note="color: #ffff00; direction: RIGHT"
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtcgactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggac tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gcttggtacc
      901 gagctcggat cgatatctgc ggcctagcta gcgcttaagg cctgttaacc ggtatggatg
      961 tatgcgtccg tcttgccctg tggctcctct ggggactcct cctgcaccag ggccagagcc
     1021 tcagccatag tcacagtgag aaggcgacag gaaccagctc gggcgccggt ttaaacgagc
     1081 atgttgcctt tggcagtgag gacatcgaga acactctggc caatatggac gacgaacaac
     1141 tggataggtt ggcctttggc gtaattcagc tcgatggtga cgggaatatc ctgctgtaca
     1201 atgctgctga aggggacatc actggcagag atcccaaaca ggtgattggg aagaacttct
     1261 tcaaggatgt tgcacctgga acggatactc ccgagtttta cggcaaattc aaggaaggcg
     1321 cagcgtcagg gaatctgaac accatgttcg aatggacgat accgacaagc aggggaccaa
     1381 ccaaggtcaa ggtgcacttg aagaaagccc tttccggagg tggcggaagt tatccttatg
     1441 acgtaccaga ctacgcactc ggaccgaaag atgaactgtg agcggccgca tagataactg
     1501 atccagtgtg ctggaattaa ttcgctgtct gcgagggcca gctgttgggg tgagtactcc
     1561 ctctcaaaag cgggcatgac ttctgcgcta agattgtcag tttccaaaaa cgaggaggat
     1621 ttgatattca cctggcccgc ggtgatgcct ttgagggtgg ccgcgtccat ctggtcagaa
     1681 aagacaatct ttttgttgtc aagcttgagg tgtggcaggc ttgagatctg gccatacact
     1741 tgagtgacaa tgacatccac tttgcctttc tctccacagg tgtccactcc caggtccaac
     1801 tgcaggtcga gcatgcatct agggcggcca attccgcccc tctccctccc ccccccctaa
     1861 cgttactggc cgaagccgct tggaataagg ccggtgtgcg tttgtctata tgttattttc
     1921 caccatattg ccgtcttttg gcaatgtgag ggcccggaaa cctggccctg tcttcttgac
     1981 gagcattcct aggggtcttt cccctctcgc caaaggaatg caaggtctgt tgaatgtcgt
     2041 gaaggaagca gttcctctgg aagcttcttg aagacaaaca acgtctgtag cgaccctttg
     2101 caggcagcgg aaccccccac ctggcgacag gtgcctctgc ggccaaaagc cacgtgtata
     2161 agatacacct gcaaaggcgg cacaacccca gtgccacgtt gtgagttgga tagttgtgga
     2221 aagagtcaaa tggctctcct caagcgtatt caacaagggg ctgaaggatg cccagaaggt
     2281 accccattgt atgggatctg atctggggcc tcggtgcaca tgctttacat gtgtttagtc
     2341 gaggttaaaa aaacgtctag gccccccgaa ccacggggac gtggttttcc tttgaaaaac
     2401 acgatgataa gcttgccaca acccgggagg cgcgccatga agttaagtcg ccagttcacc
     2461 gtgtttggca gcgcgatctt ctgcgtcgta atcttctcac tctacctgat gctggacagg
     2521 ggtcacttgg actaccctcg gggcccgcgc caggagggct cctttccgca gggccagctt
     2581 tcaatattgc aagaaaagat tgaccatttg gagcgtttgc tcgctgagaa caacgagatt
     2641 atctcaaata tcagagactc agtcatcaac ctgagcgagt ctgtggagga cggcccgcgg
     2701 gggtcaccag gcaacgccag ccaaggctcc atccacctcc actcgccaca gttggccctg
     2761 caggctgacc ccagagactg tttggatccc accggtcgcc accggaattc cgacgagaag
     2821 accactggtt ggcgaggtgg acacgttgtt gaaggactgg ctggggaact tgaacaactt
     2881 cgtgcacgac tggagcatca cccacaaggt caacgtgaac cacctgcagg tgtgagcaag
     2941 ggcgaggaga ataacatggc catcatcaag gagttcatgc gcttcaaggt gcacatggag
     3001 ggctccgtga acggccacga gttcgagatc gagggcgagg gcgagggccg cccctacgag
     3061 gcctttcaga ccgctaagct gaaggtgacc aagggtggcc ccctgccctt cgcctgggac
     3121 atcctgtccc ctcagttcat gtacggctcc aaggtctaca ttaagcaccc agccgacatc
     3181 cccgactact tcaagctgtc cttccccgag ggcttcaggt gggagcgcgt gatgaacttc
     3241 gaggacggcg gcattattca cgttaaccag gactcctccc tgcaggacgg cgtgttcatc
     3301 tacaaggtga agctgcgcgg caccaacttc ccctccgacg gccccgtaat gcagaagaag
     3361 accatgggct gggaggcctc cgaggagcgg atgtaccccg aggacggcgc cctgaagagc
     3421 gagatcaaga agaggctgaa gctgaaggac ggcggccact acgccgccga ggtcaagacc
     3481 acctacaagg ccaagaagcc cgtgcagctg cccggcgcct acatcgtcga catcaagttg
     3541 gacatcgtgt cccacaacga ggactacacc atcgtggaac agtacgaacg cgccgagggc
     3601 cgccactcca ccggcggcat ggacgagctg tacaagggcc ggccaggcgg cggctccggt
     3661 gacagatatt gggtctttgt gaaacgggtg ggcctcgagg gccaattaat taactctaga
     3721 taactgatca taatcagcca taccacattt gtagaggttt tacttgcttt aaaaaacctc
     3781 ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt taacttgttt
     3841 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
     3901 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttaacgcgtc
     3961 gagtgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac
     4021 cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt
     4081 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg
     4141 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt
     4201 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     4261 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     4321 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     4381 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     4441 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     4501 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     4561 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     4621 ttctcccttc gggaagcgtg gcgctttctc aatgctcacg ctgtaggtat ctcagttcgg
     4681 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     4741 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     4801 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     4861 tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc
     4921 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     4981 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     5041 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     5101 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     5161 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     5221 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     5281 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     5341 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     5401 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     5461 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     5521 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     5581 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     5641 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     5701 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     5761 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     5821 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     5881 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     5941 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     6001 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     6061 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     6121 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     6181 gcacatttcc ccgaaaagtg ccacctgacg tc
//
