LOCUS       sequence-76050-e        9159 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses enzymatically monobiotinylated full length RAMA 
            ectodomain.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9159)
  AUTHORS   Zenonos ZA, Rayner JC, Wright GJ
  TITLE     Towards a comprehensive Plasmodium falciparum merozoite cell surface
            and secreted recombinant protein library.
  JOURNAL   Malar J. 2014 Mar 12;13(1):93. doi: 10.1186/1475-2875-13-93.
  PUBMED    24620899
REFERENCE   2  (bases 1 to 9159)
  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: "Malar J."; 
            date: "2014-03-12"; volume: "13(1):93. doi"; pages: " 
            10.1186/1475-2875-13-93"
COMMENT     Sequence Label: RAMA-bio-His
FEATURES             Location/Qualifiers
     source          1..9159
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        7..386
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        387..590
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     541..561
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     587..611
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     primer_bind     1092..1111
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      1134..1143
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4323..4340
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    4392..4447
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4446..4465)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(6683..6701)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6769..6873
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6874..6942,6943..7734)
                     /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(7092..7111)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7905..8493
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8394..8413
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8647..8664
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8781..8802
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8817..8822,8823..8840,8841..8847)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8855..8871
                     /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     8860..8882
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8879..8895
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8879..8895
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 catgttgaca ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc
       61 atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac
      121 cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa
      181 tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag
      241 tacatcaagt gtatcatatg ccaagtccgc cccctattga cgtcaatgac ggtaaatggc
      301 ccgcctggca ttatgcccag tacatgacct tacgggactt tcctacttgg cagtacatct
      361 acgtattagt catcgctatt accatggtga tgcggttttg gcagtacacc aatgggcgtg
      421 gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt
      481 tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taataacccc gccccgttga
      541 cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct cgtttagtga
      601 accgtcagat cctcactctc ttccgcatcg ctgtctgcga gggccagctg ttgggctcgc
      661 ggttgaggac aaactcttcg cggtctttcc agtactcttg gatcggaaac ccgtcggcct
      721 ccgaacggta ctccgccacc gagggacctg agcgagtccg catcgaccgg atcggaaaac
      781 ctctcgagaa aggcgtctaa ccagtcacag tcgcaaggta ggctgagcac cgtggcgggc
      841 ggcagcgggt ggcggtcggg gttgtttctg gcggaggtgc tgctgatgat gtaattaaag
      901 taggcggtct tgagacggcg gatggtcgag gtgaggtgtg gcaggcttga gatccagctg
      961 ttggggtgag tactccctct caaaagcggg cattacttct gcgctaagat tgtcagtttc
     1021 caaaaacgag gaggatttga tattcacctg gcccgatctg gccatacact tgagtgacaa
     1081 tgacatccac tttgcctttc tctccacagg tgtccactcc caggtccaag tttgccacca
     1141 tggagtttca gacccaggta ctcatgtccc tgctgctctg catgtctggt gcggccgcat
     1201 acctggaaca gatcaagaac ggcatcagcg gccactacac cgaggaccac aacatcaaga
     1261 acaacaactg catcagcttc agcgactacg agcggagcat caagaatttc gccatcagca
     1321 gccacgccga gaacaactac gacaacatca tcaacgagta caagaagatc aaggacatca
     1381 acaacaatat caacatcctg agcagcgtgc accggaaggg ccggatcctg tacgacagct
     1441 tcctggaaat caacaagctc gagaacgaca agaaagagaa gcacgagaaa gaggacgagt
     1501 acgaggacaa cgacgagtcc tttctggaaa ccgaggaata cgaggataat gaggacgaga
     1561 agtacaacaa ggacgaggac gactacgccg agagcttcat cgagacagat gagtatgaag
     1621 ataacgagga tgacaagtat aacaaagatg aggatgatta cagcgagtcc ttcattgaga
     1681 ctgacgaata cgacgacaat gaagaggaac agtataacaa ggacgaagat gactataccg
     1741 actccttcat cgaaaccgac cactacgaga acaatgacga taagaacgag gaagaagaag
     1801 agtacaacga ccaggacaat gactacggct acaacttcct cgaaactgat gagtacgacg
     1861 atagcgaaga gtacgactat gacgacaaag agtacgggga gagtttcctc gagaaagaag
     1921 agggcgaaga gatgaaggat gaggaaatga aggacgaaga gatgaaggac gtggaaatga
     1981 aggatgaaga gatgaaggat gaagaaatga agtatgacga gatgaagaac gaggaaatga
     2041 agtacgacga aatgaaggat gaagtcatga aggacgaaga aatgaaggat gaagtgatga
     2101 aggacgaaga gatgaaggat gagcagatga agtacgagga attcaagaac gaggaatcca
     2161 agaatgaaga gtccaaaaac gaagaaagca agaatgagga aagtaaaaac gaagagttta
     2221 agaacgaaga gagtaaaaac gaggaattca aaaacgaaga gttcaaaaat gaggacatga
     2281 gctacgatga gtacatgggc tataagaaaa aagaagagga cgagagctac aataccttca
     2341 acggcgccaa gaagaacaac gccgccaata gctttctcga aaaggacctg cagggcgact
     2401 ccgacgacga gctgcacagc accttctaca gcaagaacgt ggacaaagag aactacgacg
     2461 acaagaacat cttctacggc tacagcgaca acgacgacga gagtttcctg gaaaccgaca
     2521 gctacgagga atatgaagat gaggacaagg acgtcgagga tgaatacgaa gagtccttcc
     2581 tgcagaacga cgagaagaaa atggtgttct acgacctgta caagcccgaa gagaacgagg
     2641 cctactatga gaagaagcag aagaaagaag agaaagagga aaaagaagaa aaagagcagt
     2701 ccctgaacaa gcagaacgat atggaagatc aggaagataa cgaagagtac aagtttgagg
     2761 aagagaacaa agaggacctg ctcgacgtcc agcaggacga ggaactgcct agcgagggca
     2821 agcagaaagt gaagggcaag agcttcgaca acgagcacct gaacgagatc cagaatgtgg
     2881 ccgacgtgca cgccttcatc cagaaagaca tgaagtacct ggacgacctg atcgatgagg
     2941 aacagaccat caaggacgcc gtgaagaagt ccgcctacaa gggcaacaag aagctgggca
     3001 acaacaagaa aagccagatg atcctcgaag aagaacccga ggaaaacttc gaagaggacg
     3061 ccgacgaaga actgaacaag ctgatggaac aggaaaagaa catcgtcgac aaagaaatca
     3121 agaactccaa ggccaacaag gctaacaaaa agctgcagtt caacaacgcc aacaaacaga
     3181 acaagatgta catgaagaat gagtataaca acaaggcaaa gaacaacaaa aacaacaagt
     3241 tcgagcagca gaattacgac gaatcctaca tggacgacga ttacgagcag aacgaagaat
     3301 tcaacgacaa caaccaggcc gaggacatga aggaaacaaa cgagctggac aagatcaacg
     3361 atgagctgct gaccgaccag ggccccaacg aggataccct gctggaaaac aacaacaaaa
     3421 tcttcgataa caagttcgtg gcccacaaga agcgcgagaa gtccatcagc ccccacagct
     3481 accagaaagt gtccaccaag gtgcagaaca aagaagatat ggaaaacaaa gaggaaaagc
     3541 agctgatcaa cgacgaggcc gccatgaccg ccgaggaact cgtggaactc gagaataccg
     3601 aggacgtgaa cacccctacc atggtggaaa cagaagagat cgacagcgac gagaacggca
     3661 acaaggcctc tggcgcgccg tcgacctcca tcacggccta taagagtgag ggggagtcag
     3721 cggagttctc cttcccactc aaccttggag aggaaagcct gcagggagag ttgagatgga
     3781 aggcagagaa ggctccttct tcccagtcct ggatcacctt ctccctaaag aaccaaaagg
     3841 tgtctgtgca gaagtctact agcaacccca agttccagct gtccgaaacg ctcccactca
     3901 cccttcagat accccaggtc tcccttcagt ttgctggttc tggcaacctg accctgactc
     3961 tggacagagg gatactgtat caggaagtga acctggtggt gatgaaagtg actcagcccg
     4021 acagcaacac tttgacctgt gaggtgatgg gacccacctc acccaagatg agactgatct
     4081 tgaagcagga gaatcaggag gccagggtct ccaggcagga gaaagtgatt caagtgcagg
     4141 cccctgaagc aggggtgtgg caatgtctac tgagtgaagg tgaagaggtc aagatggact
     4201 ccaagatcca ggttttatcc aaagggttga attccggcag cctgcaccac atcctggacg
     4261 cccagaagat gctgtggaac cacagggaca ggaacctgcc tcctctggcc cctctgggcc
     4321 cccaccacca tcaccaccat tgagtctaga cacgtgatta attaaggatc ccccgacctc
     4381 gacctctggc taataaagga aatttatttt cattgcaata gtgtgttgga attttttgtg
     4441 tctctcactc ggaaggacat atgggagggc aaatcatttg gtcgagatcc ctcggagatc
     4501 tctagctaga ggatcgatcc ccgccccgga cgaactaaac ctgactacga catctctgcc
     4561 ccttcttcgc ggggcagtgc atgtaatccc ttcagttggt tggtacaact tgccaactgg
     4621 gccctgttcc acatgtgaca cgggggggga ccaaacacaa aggggttctc tgactgtagt
     4681 tgacatcctt ataaatggat gtgcacattt gccaacactg agtggctttc atcctggagc
     4741 agactttgca gtctgtggac tgcaacacaa cattgccttt atgtgtaact cttggctgaa
     4801 gctcttacac caatgctggg ggacatgtac ctcccagggg cccaggaaga ctacgggagg
     4861 ctacaccaac gtcaatcaga ggggcctgtg tagctaccga taagcggacc ctcaagaggg
     4921 cattagcaat agtgtttata aggccccctt gttaacccta aacgggtagc atatgcttcc
     4981 cgggtagtag tatatactat ccagactaac cctaattcaa tagcatatgt tacccaacgg
     5041 gaagcatatg ctatcgaatt agggttagta aaagggtcct aaggaacagc gatatctccc
     5101 accccatgag ctgtcacggt tttatttaca tggggtcagg attccacgag ggtagtgaac
     5161 cattttagtc acaagggcag tggctgaaga tcaaggagcg ggcagtgaac tctcctgaat
     5221 cttcgcctgc ttcttcattc tccttcgttt agctaataga ataactgctg agttgtgaac
     5281 agtaaggtgt atgtgaggtg ctcgaaaaca aggtttcagg tgacgccccc agaataaaat
     5341 ttggacgggg ggttcagtgg tggcattgtg ctatgacacc aatataaccc tcacaaaccc
     5401 cttgggcaat aaatactagt gtaggaatga aacattctga atatctttaa caatagaaat
     5461 ccatggggtg gggacaagcc gtaaagactg gatgtccatc tcacacgaat ttatggctat
     5521 gggcaacaca taatcctagt gcaatatgat actggggtta ttaagatgtg tcccaggcag
     5581 ggaccaagac aggtgaacca tgttgttaca ctctatttgt aacaagggga aagagagtgg
     5641 acgccgacag cagcggactc cactggttgt ctctaacacc cccgaaaatt aaacggggct
     5701 ccacgccaat ggggcccata aacaaagaca agtggccact cttttttttg aaattgtgga
     5761 gtgggggcac gcgtcagccc ccacacgccg ccctgcggtt ttggactgta aaataagggt
     5821 gtaataactt ggctgattgt aaccccgcta accactgcgg tcaaaccact tgcccacaaa
     5881 accactaatg gcaccccggg gaatacctgc ataagtaggt gggcgggcca agataggggc
     5941 gcgattgctg cgatctggag gacaaattac acacacttgc gcctgagcgc caagcacagg
     6001 gttgttggtc ctcatattca cgaggtcgct gagagcacgg tgggctaatg ttgccatggg
     6061 tagcatatac tacccaaata tctggatagc atatgctatc ctaatctata tctgggtagc
     6121 ataggctatc ctaatctata tctgggtagc atatgctatc ctaatctata tctgggtagt
     6181 atatgctatc ctaatttata tctgggtagc ataggctatc ctaatctata tctgggtagc
     6241 atatgctatc ctaatctata tctgggtagt atatgctatc ctaatctgta tccgggtagc
     6301 atatgctatc ctaatagaga ttagggtagt atatgctatc ctaatttata tctgggtagc
     6361 atatactacc caaatatctg gatagcatat gctatcctaa tctatatctg ggtagcatat
     6421 gctatcctaa tctatatctg ggtagcatag gctatcctaa tctatatctg ggtagcatat
     6481 gctatcctaa tctatatctg ggtagtatat gctatcctaa tttatatctg ggtagcatag
     6541 gctatcctaa tctatatctg ggtagcatat gctatcctaa tctatatctg ggtagtatat
     6601 gctatcctaa tctgtatccg ggtagcatat gctatcctca tgataagctg tcaaacatga
     6661 gaattttctt gaagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg
     6721 ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct
     6781 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     6841 taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc
     6901 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
     6961 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc
     7021 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact
     7081 tttaaagttc tgctatgtgg cgcggtatta tcccgtgttg acgccgggca agagcaactc
     7141 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag
     7201 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
     7261 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt
     7321 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
     7381 gccataccaa acgacgagcg tgacaccacg atgcctgcag caatggcaac aacgttgcgc
     7441 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg
     7501 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
     7561 gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca
     7621 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
     7681 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca
     7741 gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     7801 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     7861 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     7921 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     7981 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     8041 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     8101 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     8161 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     8221 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     8281 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     8341 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     8401 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     8461 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     8521 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     8581 gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc
     8641 gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctctc
     8701 cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg
     8761 ggcagtgagc gcaacgcaat taatgtgagt tagctcactc attaggcacc ccaggcttta
     8821 cactttatgc ttccggctcg tatgttgtgt ggaattgtga gcggataaca atttcacaca
     8881 ggaaacagct atgaccatga ttacgccaag ctctagctag aggtcgacca attctcatgt
     8941 ttgacagctt atcatcgcag atccgggcaa cgttgttgcc attgctgcag gcgcagaact
     9001 ggtaggtatg gaagatctat acattgaatc aatattggca attagccata ttagtcattg
     9061 gttatatagc ataaatcaat attggctatt ggccattgca tacgttgtat ctatatcata
     9121 atatgtacat ttatattggc tcatgtccaa tatgaccgc
//
