LOCUS       sequence-487491-        7952 bp    DNA     circular SYN 30-SEP-2025
DEFINITION  Expression and purification of rat CAPS1 G476E mutant.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7952)
  TITLE     Martin lab plasmids
REFERENCE   2  (bases 1 to 7952)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Sep 30, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: CAPS1 G476E HA
FEATURES             Location/Qualifiers
     source          1..7952
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        196..399
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     350..370
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     475..491
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     476..492
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             4419..4445
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    4571..4692
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4608..4627)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4662..4681
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4699..5154)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4836..4857)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5048..5067
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5181..5285
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5287..5644
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5287..5478
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5310..5330)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5495..5630
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5679..6473
                     /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             5679..6473
                     /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(5733..5752)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6343..6362
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6661..6680)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6705..6752
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      7081..7669
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7570..7589
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        join(7844..7952,1..195)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg
       61 tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat
      121 tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc
      181 atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt
      241 gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac
      301 caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc
      361 ggtaggcgtg tacggtggga ggtctatata agcagagctg gtttagtgaa ccgtcagatc
      421 cgctagcgct accggactca gatctcgagc tcaagcttgg taccgggccc cccctcgagg
      481 tcgacggtat cgccaccatg ctggaccctt cgtccagcga agaggaatcg gatgagatcc
      541 tggaagagga gagcggcaag gaggtgctgg gctcggccgc gtccggagcg cgcctgtctc
      601 caagccgcac caacgagggc tcggcgggca gcgccggcat ggggggcagt ggcgccgggg
      661 ccggggtggg cgccggcggc ggtgggggta gcggcgcgag cagcggcggc ggggccgggg
      721 ggctgcagcc cagcagccgc gccggcggcg gccggccctc cagccccagc ccgtcggtgg
      781 tgagcgagaa ggagaaggaa gagttggagc ggctgcagaa ggaagaggag gagaggaaga
      841 agaggctgca gctgtacgtg ttcgtgatgc gctgcattgc ctaccccttc aacgccaagc
      901 agcccaccga catggctcgg cggcagcaga agatcagcaa acagcagctg cagactgtca
      961 aggaccgatt ccaggctttc ctcaatgggg agacccaaat cgtggcagat gaagccttta
     1021 tgaatgctgt ccagagctac tatgaggtgt ttctcaagag tgaccgtgtg gcccgcatgg
     1081 tacagagtgg gggctgctca gccaacgact ctcgggaggt cttcaagaaa cacatcgaga
     1141 agagggtacg cagcctacct gagattgatg gcctcagcaa ggagactgtg ctaagctcct
     1201 ggatggctaa gtttgacgcc atctaccgtg gtgaggagga ccccaggaag cagcaagccc
     1261 ggatgacagc cagtgcagct tctgagttga ttctgagcaa ggaacagctc tatgagatgt
     1321 tccagaacat tcttgggatc aagaaatttg aacatcaact cctatataat gcgtgtcagc
     1381 tggacaatcc agatgagcag gcagcccaga tcagacgaga gctggacgga cgtctccaaa
     1441 tggcagacca aatagccagg gaacgcaaat ttcctaagtt cgtatccaaa gaaatggaaa
     1501 atatgtacat cgaggagctg aagtcctccg tcaacctact catggccaac ttggagagca
     1561 tgccggtgtc caagggtgga gagttcaagc tgcagaaact gaaacgcagt cacaacgcgt
     1621 ccatcattga catgggtgag gagagcgaga accagctgtc caagtcggat gtcctgctgt
     1681 ccttctcctt ggaggttgtc atcatggaag tccaaggcct taagtccttg gctccaaacc
     1741 gaatcgtgta ctgcacaatg gaggtggagg gtggagagaa acttcagact gatcaagcag
     1801 aggcttctaa accaacctgg ggcactcaag gcgacttctc tacaacccac gcgctgccag
     1861 ctgttaaagt aaagctattc acagagagca ctggtgtcct ggccttggaa gacaaggaac
     1921 tcgagcgggt tatcctccat cccaccccaa acagccccaa acaatccgag tggcacaaaa
     1981 tgacagtctc caaaaactgc cccgatcaag acctcaaaat caaactcgct gtccgaatgg
     2041 ataagcctca aaacatgaag cactctgggt acttatggac cattggtaag aatgtgtgga
     2101 agaggtggaa gaaaaggttc tttgtgttgg tgcaggtcag ccagtacact tttgccatgt
     2161 gcagctatcg agaaaagaaa gcagaacctc aggaactcct acagctggat ggctacaccg
     2221 tggattacac cgaccctcag ccaggtttgg agggtggccg agccttcttc aatgcagtca
     2281 aagaaggaga cacggtgata tttgccagtg acgatgagca agatcgaatc ctgtgggtcc
     2341 aagctatgta cagggccacc gggcagtcgc acaaacctgt gccccctacc caggtgcaga
     2401 agctcaacgc caagggcgga aacgtgcctc agctggatgc ccccatctcc cagttttacg
     2461 cagatagagc tcaaaagcat ggcatggatg aatttatctc gtccaacccc tgtaactttg
     2521 accacgcttc cctctttgag atggtacagc ggctgacttt ggatcacaga ctaaacgatt
     2581 cctattcttg cctgggctgg ttcagtcctg gccaggtgtt tgtgttagac gagtactgcg
     2641 ctcggaatgg agtccgagga tgtcatcggc atctctgtta ccttagagac ttgctggaac
     2701 gggcagaaaa tggcgccatg atcgacccca cgcttcttca ttacagcttt gccttctgcg
     2761 catcccatgt ccacgggaac aggcctgatg gaatcggaac tgtgactgtt gaagaaaaag
     2821 agcgctttga agaaatcaaa gagagactcc gagttctgct ggaaaatcaa attacacatt
     2881 tcaggtactg ctttccgttt ggtcgacctg aaggtgcttt gaaagcgact ctctcacttt
     2941 tggaacgggt tttgatgaaa gatattgtca ctccagtacc ccaagaggag gtcaaaactg
     3001 tcatccgtaa atgcctagag caggctgctc tagtcaacta ctctcgccta tctgagtatg
     3061 ccaaaatcga agaaaatgta ggccgcttaa tcactcccgc caaaaagctc gaagacacaa
     3121 tccgtcttgc tgaactagtc attgaagtgc tgcaacaaaa cgaagagcac cacgccgagg
     3181 cctttgcgtg gtggtcagac ttaatggtgg agcacgcgga gacgttcctg tcactctttg
     3241 cagtggacat ggatgcagca ttagaggtgc aacccccaga cacgtgggac agtttcccac
     3301 tcttccagct gctgaatgac tttctccgta cagactataa cttgtgcaat ggaaagtttc
     3361 acaaacacct gcaggatctg ttcgccccac tcgttgttag atacgtggat ctgatggagt
     3421 cctcaattgc acaatctatc cacaggggct ttgagcggga gtcatgggaa ccagtcaata
     3481 acggatcggg tacatcagaa gatttgttct ggaaacttga tgcccttcag actttcatcc
     3541 gtgacctaca ctggcctgag gaagagtttg gaaagcactt agagcagcgg ctaaaactaa
     3601 tggcaagcga catgattgag tcctgcgtca agagaaccag gattgcattt gaagttaagc
     3661 tgcaaaaaac cagtcgatca acagattttc gagtcccaca gtcaatatgc accatgttta
     3721 atgttatggt tgatgccaaa gctcaatcaa caaaactttg cagcatggaa atgggccagg
     3781 agcatcaata ccattcaaaa atagacgaac taattgaaga aaccgttaag gaaatgatca
     3841 cactcttggt ggcaaagttt gtaactatat tggaaggcgt actggcaaag ttatccagat
     3901 acgacgaggg gactttgttt tcttcgtttc tgtcatttac ggtgaaggcg gcatctaagt
     3961 atgtggacgt acctaaaccc gggatggatg tggccgacgc ctacgtgact ttcgtccgcc
     4021 actctcagga tgtccttcgt gataaggtca atgaggagat gtatatagaa aggttatttg
     4081 atcaatggta caacagctcc atgaacgtca tctgcacctg gctgaccgac cggatggacc
     4141 tccagcttca catttatcag ctgaaaacac taattaggat ggtaaagaaa acctacagag
     4201 atttccgatt gcaaggggtc ctggattcca ccttaaacag caagacctat gagaccattc
     4261 gaaatcggct cactgtagag gaagctaccg cgtccgtgag tgagggaggg ggcttgcaag
     4321 gcatcagcat gaaggacagc gatgaagagg acgaggagga tgacggtgga ggaggttctg
     4381 gccaccacac tggactagtg gatccaccgg tcgccaccta cccatacgat gttccagatt
     4441 acgcttaaag cggccgcgac tctagatcat aatcagccat accacatttg tagaggtttt
     4501 acttgcttta aaaaacctcc cacacctccc cctgaacctg aaacataaaa tgaatgcaat
     4561 tgttgttgtt aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     4621 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     4681 caatgtatct taaggcgtaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt
     4741 tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat
     4801 caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat
     4861 taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac
     4921 tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc
     4981 ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga
     5041 gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca
     5101 cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtcaggtg
     5161 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     5221 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     5281 agagtcctga ggcggaaaga accagctgtg gaatgtgtgt cagttagggt gtggaaagtc
     5341 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag
     5401 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     5461 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
     5521 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
     5581 ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
     5641 caaagatcga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc
     5701 acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga
     5761 caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt
     5821 ttgtcaagac cgacctgtcc ggtgccctga atgaactgca agacgaggca gcgcggctat
     5881 cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg
     5941 gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg
     6001 ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc
     6061 cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga
     6121 tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag
     6181 ccgaactgtt cgccaggctc aaggcgagca tgcccgacgg cgaggatctc gtcgtgaccc
     6241 atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg
     6301 actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata
     6361 ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg
     6421 ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac
     6481 tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc
     6541 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat
     6601 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccctaggg ggaggctaac
     6661 tgaaacacgg aaggagacaa taccggaagg aacccgcgct atgacggcaa taaaaagaca
     6721 gaataaaacg cacggtgttg ggtcgtttgt tcataaacgc ggggttcggt cccagggctg
     6781 gcactctgtc gataccccac cgagacccca ttggggccaa tacgcccgcg tttcttcctt
     6841 ttccccaccc caccccccaa gttcgggtga aggcccaggg ctcgcagcca acgtcggggc
     6901 ggcaggccct gccatagcct caggttactc atatatactt tagattgatt taaaacttca
     6961 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     7021 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     7081 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     7141 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     7201 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     7261 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     7321 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     7381 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     7441 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     7501 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     7561 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     7621 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     7681 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
     7741 gttatcccct gattctgtgg ataaccgtat taccgccatg cattagttat taatagtaat
     7801 caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg
     7861 taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt
     7921 atgttcccat agtaacgcca atagggactt tc
//
