LOCUS       sequence-308752-        3971 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cell-free expression vector for Zika virus prM protein with C-term 
            mCherry.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3971)
  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 3971)
  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_ZIKV PrM
FEATURES             Location/Qualifiers
     source          1..3971
                     /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(692..716)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             718..741
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     primer_bind     complement(878..896)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1146..1165
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             1450..1473
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     terminator      1696..1742
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(1820..1836)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1820..1836)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1833..1855)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1844..1860
                     /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(1868..1874,1875..1892,1893..1898))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1913..1934
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2051..2068)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2222..2810)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2302..2321)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2981..3772,3773..3841))
                     /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     3604..3623
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3842..3946)
                     /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 ggcttcgcag agatcactag acgcgggagt gcatactaca tgtacttgga taggagcgat
      241 gccgggaagg ccatttcgtt tgctaccaca ttgggagtga acaagtgcca cgtacagatc
      301 atggacctcg ggcacatgtg tgacgccacc atgagttatg agtgccctat gctggatgag
      361 ggagtggaac cagatgatgt cgattgctgg tgcaacacga catcaacttg ggttgtgtac
      421 ggaacctgtc atcacaaaaa aggtgaggca cggcgatcta gaagagccgt gacgctccct
      481 tctcactcta caaggaagtt gcaaacgcgg tcgcagacct ggttagaatc aagagaatac
      541 acgaagcact tgatcaaggt tgaaaactgg atattcagga accccgggtt tgcgctagtg
      601 gccgttgcca ttgcctggct tttgggaagc tcgacgagcc aaaaagtcat atacttggtc
      661 atgatactgc tgattgcccc ggcatacagt aacccagctt tcttgtacaa agtggtgctg
      721 gaggtgctct tccagggccc gaagggcgag gaggataaca tggccatcat caaggagttc
      781 atgcgcttca aggtgcacat ggagggctcc gtgaacggcc acgagttcga gatcgagggc
      841 gagggcgagg gccgccccta cgagggcacc cagaccgcca agctgaaggt gaccaagggt
      901 ggccccctgc ccttcgcctg ggacatcctg tcccctcagt tcatgtacgg ctccaaggcc
      961 tacgtgaagc accccgccga catccccgac tacttgaagc tgtccttccc cgagggcttc
     1021 aagtgggagc gcgtgatgaa cttcgaggac ggcggcgtgg tgaccgtgac ccaggactcc
     1081 tccctgcagg acggcgagtt catctacaag gtgaagctgc gcggcaccaa cttcccctcc
     1141 gacggccccg taatgcagaa gaagactatg ggctgggagg cctcctccga gcggatgtac
     1201 cccgaggacg gcgccctgaa gggcgagatc aagcagaggc tgaagctgaa ggacggcggc
     1261 cactacgacg ctgaggtcaa gaccacctac aaggccaaga agcccgtgca gctgcccggc
     1321 gcctacaacg tcaacatcaa gttggacatc acctcccaca acgaggacta caccatcgtg
     1381 gaacagtacg aacgcgccga gggccgccac tccaccggcg gcatggacga gctgtacaag
     1441 acctctggtc accaccatca ccaccatcac cactaagcgg ccgccctcct cctcctttct
     1501 tgttcctttc acgtcgcctt ctcggttgta gctggcagac gacgagtctt acttttacgt
     1561 gtacttctct atagatgatg tatgatctct ctgcatgcgt gttcgtgcat gtgtccgtgt
     1621 gttgggtacg cgtggtaccc tgcaggaagg aagctgagtt ggctgctgcc accgctgagc
     1681 aataactagt aattactagc ataacccctt ggggcctcta aacgggtctt gagggggttt
     1741 tttgctgaaa ggaggacagc tgatgattgt catgcttgcc atctgttttc ttgcaaggtc
     1801 agaggaattc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
     1861 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
     1921 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     1981 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     2041 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     2101 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     2161 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     2221 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     2281 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     2341 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     2401 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     2461 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     2521 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     2581 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     2641 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     2701 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     2761 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaggatcctt
     2821 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     2881 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     2941 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     3001 aggcacctat ctcagcgatc tgtctagttc gttcatccat agttgcctga ctccccgtcg
     3061 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     3121 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     3181 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     3241 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     3301 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     3361 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     3421 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     3481 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     3541 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     3601 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     3661 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     3721 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     3781 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     3841 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     3901 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     3961 aagtgccacc t
//
