LOCUS       sequence-305924-        3779 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cell-free expression vector for Zika virus Capsid protein with 
            C-term mCherry.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3779)
  AUTHORS   Varasteh Moradi S, Gagoski D, Mureev S, Walden P, McMahon KA, Parton
            RG, Johnston WA, Alexandrov K
  TITLE     Mapping Interactions among Cell-Free Expressed Zika Virus Proteins.
  JOURNAL   J Proteome Res. 2020 Apr 3;19(4):1522-1532. doi: 
            10.1021/acs.jproteome.9b00771. Epub 2020 Mar 9.
  PUBMED    32081002
REFERENCE   2  (bases 1 to 3779)
  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"; doi: 
            "10.1021/acs.jproteome.9b00771"; journalName: "J Proteome Res"; 
            date: "2020-04-3- 3"; volume: "19"; issue: "4"; pages: "1522-1532"
COMMENT     Sequence Label: pCellFree_G06_ZIKVC
FEATURES             Location/Qualifiers
     source          1..3779
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     7..26
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        7..25
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    160..184
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(500..524)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             526..549
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     primer_bind     complement(686..704)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     954..973
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             1258..1281
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     terminator      1504..1550
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(1628..1644)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1628..1644)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1641..1663)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1652..1668
                     /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)."
     promoter        complement(join(1676..1682,1683..1700,1701..1706))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1721..1742
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1859..1876)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2030..2618)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2110..2129)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2789..3580,3581..3649))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERTRQIAEIGAS
                     LIKHW"
     primer_bind     3412..3431
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3650..3754)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacgtctaat acgactcact atagggacat cttaagttta ttttatttta ttttatttta
       61 ttttatttta ttttatttta ttttatttta ttttatttaa ccatgacagt aatgtataaa
      121 gtctgtaaag acattaaaca cgtaagtgaa accatgggca caagtttgta caaaaaagca
      181 ggcttcatga aaaaccccaa agaagaaatc cggaggatcc ggattgtcaa tatgctaaaa
      241 cgcggagtag cccgtgtaaa ccccttggga ggtttgaaga ggttgccagc cggacttctg
      301 ctgggtcatg gacccatcag aatggttttg gcgatactag cctttttgag atttacagca
      361 atcaagccat cactgggcct tatcaacaga tggggttccg tggggaaaaa agaggctatg
      421 gaaataataa agaagttcaa gaaagatctt gctgccatgt tgagaataat caatgctagg
      481 aaagagagga agagacgtaa cccagctttc ttgtacaaag tggtgctgga ggtgctcttc
      541 cagggcccga agggcgagga ggataacatg gccatcatca aggagttcat gcgcttcaag
      601 gtgcacatgg agggctccgt gaacggccac gagttcgaga tcgagggcga gggcgagggc
      661 cgcccctacg agggcaccca gaccgccaag ctgaaggtga ccaagggtgg ccccctgccc
      721 ttcgcctggg acatcctgtc ccctcagttc atgtacggct ccaaggccta cgtgaagcac
      781 cccgccgaca tccccgacta cttgaagctg tccttccccg agggcttcaa gtgggagcgc
      841 gtgatgaact tcgaggacgg cggcgtggtg accgtgaccc aggactcctc cctgcaggac
      901 ggcgagttca tctacaaggt gaagctgcgc ggcaccaact tcccctccga cggccccgta
      961 atgcagaaga agactatggg ctgggaggcc tcctccgagc ggatgtaccc cgaggacggc
     1021 gccctgaagg gcgagatcaa gcagaggctg aagctgaagg acggcggcca ctacgacgct
     1081 gaggtcaaga ccacctacaa ggccaagaag cccgtgcagc tgcccggcgc ctacaacgtc
     1141 aacatcaagt tggacatcac ctcccacaac gaggactaca ccatcgtgga acagtacgaa
     1201 cgcgccgagg gccgccactc caccggcggc atggacgagc tgtacaagac ctctggtcac
     1261 caccatcacc accatcacca ctaagcggcc gccctcctcc tcctttcttg ttcctttcac
     1321 gtcgccttct cggttgtagc tggcagacga cgagtcttac ttttacgtgt acttctctat
     1381 agatgatgta tgatctctct gcatgcgtgt tcgtgcatgt gtccgtgtgt tgggtacgcg
     1441 tggtaccctg caggaaggaa gctgagttgg ctgctgccac cgctgagcaa taactagtaa
     1501 ttactagcat aaccccttgg ggcctctaaa cgggtcttga gggggttttt tgctgaaagg
     1561 aggacagctg atgattgtca tgcttgccat ctgttttctt gcaaggtcag aggaattcgt
     1621 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
     1681 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
     1741 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     1801 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     1861 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     1921 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     1981 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     2041 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     2101 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     2161 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     2221 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     2281 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     2341 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     2401 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     2461 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     2521 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     2581 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga ggatcctttg atcttttcta
     2641 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     2701 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     2761 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     2821 cagcgatctg tctagttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     2881 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     2941 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     3001 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     3061 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     3121 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     3181 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     3241 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     3301 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     3361 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     3421 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     3481 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     3541 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     3601 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     3661 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     3721 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacct
//
