LOCUS       sequence-120644-        7665 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Expresses full-length P. vivax CyRPA ectodomain in mammalian cells. 
            C-terminal rat Cd4d3+4 tag, biotinylation sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7665)
  AUTHORS   Hostetler JB, Sharma S, Bartholdson SJ, Wright GJ, Fairhurst RM, 
            Rayner JC
  TITLE     A Library of Plasmodium vivax Recombinant Merozoite Proteins Reveals
            New Vaccine Candidates and Protein-Protein Interactions.
  JOURNAL   PLoS Negl Trop Dis. 2015 Dec 23;9(12):e0004264. doi: 
            10.1371/journal.pntd.0004264. eCollection 2015 Dec.
  PUBMED    26701602
REFERENCE   2  (bases 1 to 7665)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "PLoS Negl 
            Trop Dis."; date: "2015-12-23"; volume: "9(12):e0004264. doi"; 
            pages: " 10.1371/journal.pntd.0004264. eCollection 2015 Dec"
COMMENT     Sequence Label: CyRPA-bio
FEATURES             Location/Qualifiers
     source          1..7665
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        154..533
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        534..737
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     688..708
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     734..758
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     primer_bind     1239..1258
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      1281..1290
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    3045..3100
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3099..3118)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5336..5354)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5422..5526
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5527..5595,5596..6387)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(5745..5764)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6558..7146
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7047..7066
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7300..7317
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7434..7455
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7470..7475,7476..7493,7494..7500)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7508..7524
                     /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)."
     primer_bind     7513..7535
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7532..7548
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7532..7548
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 agatctatac attgaatcaa tattggcaat tagccatatt agtcattggt tatatagcat
       61 aaatcaatat tggctattgg ccattgcata cgttgtatct atatcataat atgtacattt
      121 atattggctc atgtccaata tgaccgccat gttgacattg attattgact agttattaat
      181 agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac
      241 ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa
      301 tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt
      361 atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca agtccgcccc
      421 ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttac
      481 gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc atggtgatgc
      541 ggttttggca gtacaccaat gggcgtggat agcggtttga ctcacgggga tttccaagtc
      601 tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg gactttccaa
      661 aatgtcgtaa taaccccgcc ccgttgacgc aaatgggcgg taggcgtgta cggtgggagg
      721 tctatataag cagagctcgt ttagtgaacc gtcagatcct cactctcttc cgcatcgctg
      781 tctgcgaggg ccagctgttg ggctcgcggt tgaggacaaa ctcttcgcgg tctttccagt
      841 actcttggat cggaaacccg tcggcctccg aacggtactc cgccaccgag ggacctgagc
      901 gagtccgcat cgaccggatc ggaaaacctc tcgagaaagg cgtctaacca gtcacagtcg
      961 caaggtaggc tgagcaccgt ggcgggcggc agcgggtggc ggtcggggtt gtttctggcg
     1021 gaggtgctgc tgatgatgta attaaagtag gcggtcttga gacggcggat ggtcgaggtg
     1081 aggtgtggca ggcttgagat ccagctgttg gggtgagtac tccctctcaa aagcgggcat
     1141 tacttctgcg ctaagattgt cagtttccaa aaacgaggag gatttgatat tcacctggcc
     1201 cgatctggcc atacacttga gtgacaatga catccacttt gcctttctct ccacaggtgt
     1261 ccactcccag gtccaagttt gccaccatgg agtttcagac ccaggtactc atgtccctgc
     1321 tgctctgcat gtctggtgcg gccgccacaa atacccagag caagaacatc atcatcctga
     1381 acgacgagat caccaccatc aagagcccca tccactgtat taccgatatc tacttcctgt
     1441 tccgcaacga gctgtacaag acctgcatcc agcacgtgat caagggccgg accgagatcc
     1501 atgtgctggt gcagaagaag atcaacagcg cctgggagac acagaccacc ctgttcaagg
     1561 atcacatgtg gttcgagctg cccagcgtgt tcaacttcat ccacaacgat gagatcatca
     1621 ttgtgatctg ccggtacaag cagcggagca agagagaggg caccatctgc aagcggtgga
     1681 acagcgtgac cggcacaatc taccagaaag aagatgtgca gatcgacaaa gaggccttcg
     1741 ccaacaagaa cctggaaagc taccagagcg tgcccctgac cgtgaagaac aagaagttcc
     1801 tgctgatctg cggcatcctg agctacgagt acaagacagc caacaaggac aatttcatca
     1861 gctgcgtggc cagcgaggac aagggcagaa cctggggcac caagatcctg atcaactacg
     1921 aggaactgca gaaaggcgtg ccctacttct acctgcggcc catcatcttc ggcgacgagt
     1981 tcggcttcta cttttacagc cggatcagca ccaacaacgc cgccagaggc ggcaactaca
     2041 tgacctgcac cctggacgtg accaacgagg gcaagaaaga gtacaagttc aagtgcaagc
     2101 acgtgtccct gatcaagccc gacaagagcc tgcagaacgt ggccaagctg aacggctact
     2161 acatcaccag ctacgtgaag aaggacaact tcaacgagtg ctacctgtac tacaccgagc
     2221 agaacgccat cgtcgtgaag cccaaggtgc agaacgacga cctgaatggc tgctacggcg
     2281 gcagcttcgt gaagctggat gagagcaagg ccctgttcat ctacagcacc ggctatggcg
     2341 tgcagaacat ccacaccctg tactatacca gatacgacgg cgcgccgtcg acctccatca
     2401 cggcctataa gagtgagggg gagtcagcgg agttctcctt cccactcaac cttggagagg
     2461 aaagcctgca gggagagttg agatggaagg cagagaaggc tccttcttcc cagtcctgga
     2521 tcaccttctc cctaaagaac caaaaggtgt ctgtgcagaa gtctactagc aaccccaagt
     2581 tccagctgtc cgaaacgctc ccactcaccc ttcagatacc ccaggtctcc cttcagtttg
     2641 ctggttctgg caacctgacc ctgactctgg acagagggat actgtatcag gaagtgaacc
     2701 tggtggtgat gaaagtgact cagcccgaca gcaacacttt gacctgtgag gtgatgggac
     2761 ccacctcacc caagatgaga ctgatcttga agcaggagaa tcaggaggcc agggtctcca
     2821 ggcaggagaa agtgattcaa gtgcaggccc ctgaagcagg ggtgtggcaa tgtctactga
     2881 gtgaaggtga agaggtcaag atggactcca agatccaggt tttatccaaa gggttgaatt
     2941 ccggatcact gcatcatatt ctggatgcac agaaaatggt gtggaatcat cgttaagtct
     3001 agacacgtga ttaattaagg atcccccgac ctcgacctct ggctaataaa ggaaatttat
     3061 tttcattgca atagtgtgtt ggaatttttt gtgtctctca ctcggaagga catatgggag
     3121 ggcaaatcat ttggtcgaga tccctcggag atctctagct agaggatcga tccccgcccc
     3181 ggacgaacta aacctgacta cgacatctct gccccttctt cgcggggcag tgcatgtaat
     3241 cccttcagtt ggttggtaca acttgccaac tgggccctgt tccacatgtg acacgggggg
     3301 ggaccaaaca caaaggggtt ctctgactgt agttgacatc cttataaatg gatgtgcaca
     3361 tttgccaaca ctgagtggct ttcatcctgg agcagacttt gcagtctgtg gactgcaaca
     3421 caacattgcc tttatgtgta actcttggct gaagctctta caccaatgct gggggacatg
     3481 tacctcccag gggcccagga agactacggg aggctacacc aacgtcaatc agaggggcct
     3541 gtgtagctac cgataagcgg accctcaaga gggcattagc aatagtgttt ataaggcccc
     3601 cttgttaacc ctaaacgggt agcatatgct tcccgggtag tagtatatac tatccagact
     3661 aaccctaatt caatagcata tgttacccaa cgggaagcat atgctatcga attagggtta
     3721 gtaaaagggt cctaaggaac agcgatatct cccaccccat gagctgtcac ggttttattt
     3781 acatggggtc aggattccac gagggtagtg aaccatttta gtcacaaggg cagtggctga
     3841 agatcaagga gcgggcagtg aactctcctg aatcttcgcc tgcttcttca ttctccttcg
     3901 tttagctaat agaataactg ctgagttgtg aacagtaagg tgtatgtgag gtgctcgaaa
     3961 acaaggtttc aggtgacgcc cccagaataa aatttggacg gggggttcag tggtggcatt
     4021 gtgctatgac accaatataa ccctcacaaa ccccttgggc aataaatact agtgtaggaa
     4081 tgaaacattc tgaatatctt taacaataga aatccatggg gtggggacaa gccgtaaaga
     4141 ctggatgtcc atctcacacg aatttatggc tatgggcaac acataatcct agtgcaatat
     4201 gatactgggg ttattaagat gtgtcccagg cagggaccaa gacaggtgaa ccatgttgtt
     4261 acactctatt tgtaacaagg ggaaagagag tggacgccga cagcagcgga ctccactggt
     4321 tgtctctaac acccccgaaa attaaacggg gctccacgcc aatggggccc ataaacaaag
     4381 acaagtggcc actctttttt ttgaaattgt ggagtggggg cacgcgtcag cccccacacg
     4441 ccgccctgcg gttttggact gtaaaataag ggtgtaataa cttggctgat tgtaaccccg
     4501 ctaaccactg cggtcaaacc acttgcccac aaaaccacta atggcacccc ggggaatacc
     4561 tgcataagta ggtgggcggg ccaagatagg ggcgcgattg ctgcgatctg gaggacaaat
     4621 tacacacact tgcgcctgag cgccaagcac agggttgttg gtcctcatat tcacgaggtc
     4681 gctgagagca cggtgggcta atgttgccat gggtagcata tactacccaa atatctggat
     4741 agcatatgct atcctaatct atatctgggt agcataggct atcctaatct atatctgggt
     4801 agcatatgct atcctaatct atatctgggt agtatatgct atcctaattt atatctgggt
     4861 agcataggct atcctaatct atatctgggt agcatatgct atcctaatct atatctgggt
     4921 agtatatgct atcctaatct gtatccgggt agcatatgct atcctaatag agattagggt
     4981 agtatatgct atcctaattt atatctgggt agcatatact acccaaatat ctggatagca
     5041 tatgctatcc taatctatat ctgggtagca tatgctatcc taatctatat ctgggtagca
     5101 taggctatcc taatctatat ctgggtagca tatgctatcc taatctatat ctgggtagta
     5161 tatgctatcc taatttatat ctgggtagca taggctatcc taatctatat ctgggtagca
     5221 tatgctatcc taatctatat ctgggtagta tatgctatcc taatctgtat ccgggtagca
     5281 tatgctatcc tcatgataag ctgtcaaaca tgagaatttt cttgaagacg aaagggcctc
     5341 gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt
     5401 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
     5461 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
     5521 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc
     5581 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
     5641 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt
     5701 cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta
     5761 ttatcccgtg ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat
     5821 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga
     5881 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca
     5941 acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact
     6001 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc
     6061 acgatgcctg cagcaatggc aacaacgttg cgcaaactat taactggcga actacttact
     6121 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt
     6181 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt
     6241 gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt
     6301 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
     6361 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag
     6421 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     6481 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     6541 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     6601 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     6661 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     6721 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     6781 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     6841 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     6901 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     6961 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     7021 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     7081 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     7141 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     7201 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag
     7261 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa
     7321 gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc
     7381 agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg
     7441 agttagctca ctcattaggc accccaggct ttacacttta tgcttccggc tcgtatgttg
     7501 tgtggaattg tgagcggata acaatttcac acaggaaaca gctatgacca tgattacgcc
     7561 aagctctagc tagaggtcga ccaattctca tgtttgacag cttatcatcg cagatccggg
     7621 caacgttgtt gccattgctg caggcgcaga actggtaggt atgga
//
