LOCUS       sequence-396580-        6216 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Mammalian cell expression of Nipah virus fusion protein with 
            prefusion stabilizing mutations.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6216)
  TITLE     Structures of virus fusion protein ectodomain in pre and post fusion
            conformation
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6216)
  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: 
            "Unpublished"
COMMENT     Sequence Label: p-alpha-H-NiVop08-Foldon
FEATURES             Location/Qualifiers
     source          1..6216
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        5..384
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        386..661
                     /label=chicken beta-actin promoter
     intron          662..1678
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1602..1624
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     regulatory      1760..1769
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3326..3349
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             3353..3376
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     CDS             3383..3406
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             3446..3469
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(3571..3590)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3636..3691
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3690..3709)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4052..4068)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4052..4068)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4065..4087)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4076..4092
                     /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(4100..4106,4107..4124,4125..4130))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4145..4166
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4295..4312)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4466..5054)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4546..4565)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5225..6016,6017..6085))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     5848..5867
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6086..6190)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ggtcgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat
       61 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg
      121 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
      181 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
      241 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
      301 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
      361 gtattagtca tcgctattac catggtcgag gtgagcccca cgttctgctt cactctcccc
      421 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca
      481 gcgatggggg cggggggggg gggggggcgc gccagggggg gggggggggg gsgagggggg
      541 gggsggggsg aggcggagag gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt
      601 tccttttatg gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc
      661 gggagtcgct gcgcgctgcc ttcgccccgt gccccgctcc gccgccgcct cgcgccgccc
      721 gccccggctc tgactgaccg cgttactccc acaggtgagc gggcgggacg gcccttctcc
      781 tccgggctgt aattagcgct tggtttaatg acggcttgtt tcttttctgt ggctgcgtga
      841 aagccttgag gggctccggg agggcccttt gtgcgggggg agcggctcgg ggggtgcgtg
      901 cgtgtgtgtg tgcgtgggga gcgccgcgtg cggctccgcg ctgcccggcg gctgtgagcg
      961 ctgcgggcgc ggcgcggggc tttgtgcgct ccgcagtgtg cgcgagggga gcgcggccgg
     1021 gggcggtgcc ccgcggtgcg gggggggctg cgaggggaac aaaggctgcg tgcggggtgt
     1081 gtgcgtgggg gggtgagcag ggggtgtggg cgcgtcggtc gggctgcaac cccccctgca
     1141 cccccctccc cgagttgctg agcacggccc ggcttcgggt gcggggctcc gtacggggcg
     1201 tggcgcgggg ctcgccgtgc cgggcggggg gtggcggcag gkgggggtgc cgggcggggc
     1261 ggggccgcct cgggccgggg rgggctcggg ggaggggcgc ggcggccccc ggagcgccgg
     1321 cggctgtcga ggcgcggcga gccgcagcca ttgcctttta tggtaatcgt gcgagagggc
     1381 gcagggactt cctttgtccc aaatctgtgc ggagccgaaa tctgggaggc gccgccgcac
     1441 cccctctagc gggcgcgggg cgaagcggtg cggcgccggc aggaaggaaa tgggcgggga
     1501 gggccttcgt gcgtcgccgc gccgccgtcc ccttctccct ctccagcctc ggggctgtcc
     1561 gcggggggac ggctgccttc gggggggacg gggcagggcg gggttcggct tctggcgtgt
     1621 gaccggcggc tctagcgcct ctgctaacca tgttcatgcc ttcttctttt tcctacagct
     1681 cctgggcaac gtgctggttg ttgtgctgtc tcatcatttt ggcaaagaat tgcggccgtc
     1741 tcaggccgag ttcggtaccg ccaccatggt ggtgatcctg gacaagaggt gctactgcaa
     1801 cctgctgatc ctgatcctga tgatcagcga gtgcagcgtg ggcatcctgc actacgagaa
     1861 gctgagcaag atcggcctgg tgaagggcgt gaccaggaag tacaagatca agagcaaccc
     1921 cctgaccaag gacatcgtga tcaagatgat ccccaacgtg agcaacatga gccagtgcac
     1981 cggcagcgtg atggagaact acaagaccag gctgaacggc atcctgaccc ccatcaaggg
     2041 cgccctggag atctacaaga acaacaccca cgactgcgtg ggcgacgtga ggctggccgg
     2101 cgtgtgcatg gccggcgtgg ccatcggcat cgccaccgcc gcccagatca ccgccggcgt
     2161 ggccctgtac gaggccatga agaacgccga caacatcaac aagctgaaga gcagcatcga
     2221 gagcaccaac gaggccgtgg tgaagctgca ggagaccgcc gagaagaccg tgtacgtgtt
     2281 caccgccctg caggactaca tcaacaccaa cctggtgccc accatcgaca agatcccctg
     2341 caagcagacc gagctgagcc tggacctggc cctgagcaag tacctgagcg acctgctgtt
     2401 cgtgttcggc cccaacctgc aggaccccgt gagcaacagc atgaccatcc aggccatcag
     2461 ccaggccttc ggcggcaact acgagaccct gctgaggacc ctgggctacg ccaccgagga
     2521 cttcgacgac ctgctggaga gcgacagcat caccggccag atcatctacg tggacctgag
     2581 cagctactac atcatcgtga gggtgtactt ccccatcctg accgagatcc agcaggccta
     2641 catccaggag ctgctgcccg tgagcttcaa caacgacaac agcgagtgga tcagcatcgt
     2701 gcccaacttc atcctggtga ggaacaccct gatcagcaac atcgagatcg gcttctgcct
     2761 gatcaccaag aggagcgtga tctgcaacca ggactacgcc acccccatga ccaacaacat
     2821 gagggagtgc ctgaccggca gcaccgagaa gtgccccagg gagctggtgg tgagcagcca
     2881 cgtgcccagg ttcgccctga gcaacggcgt gctgttcgcc aactgcatca gcgtgacctg
     2941 ccagtgccag accaccggca gggccatcag ccagagcggc gagcagaccc tgctgatgat
     3001 cgacaacacc acctgcccca ccgccgtgct gggcaacgtg atcatcagcc tgggcaagta
     3061 cctgggcagc gtgaactaca acagcgaggg catcgccatc ggcccccccg tgttcaccga
     3121 caaggtggac atcagcagcc agatcagcag catgaaccag agcctgcagc agagcaagga
     3181 ctacatcaag gaggcccaga ggctgctgga caccgtgaac cccagcctgg gatccggata
     3241 catccccgag gcccccagag atggccaggc ctacgtgcgg aaggacggcg agtgggtact
     3301 gctgagcaca ttcctgggca gatccctgga ggtgctgttc cagggcccag gccatcacca
     3361 ccatcaccac catcatagcg cctggtccca cccccagttc gagaagggcg gcggtagtgg
     3421 agggggcgga tctggcggct cagcttggag ccacccccag ttcgaaaagt gataatgact
     3481 cgagcgataa ttcactcctc aggtgcaggc tgcctatcag aaggtggtgg ctggtgtggc
     3541 caatgccctg gctcacaaat accactgaga tctttttccc tctgccaaaa attatgggga
     3601 catcatgaag ccccttgagc atctgacttc tggctaataa aggaaattta ttttcattgc
     3661 aatagtgtgt tggaattttt tgtgtctctc actcggaagg acatatggga gggcaaatca
     3721 tttaaaacat cagaatgagt atttggttta gagtttggca acatatgccc atatgctggc
     3781 tgccatgaac aaaggttggc tataaagagg tcatcagtat atgaaacagc cccctgctgt
     3841 ccattcctta ttccatagaa aagccttgac ttgaggttag atttttttta tattttgttt
     3901 tgtgttattt ttttctttaa catccctaaa attttcctta catgttttac tagccagatt
     3961 tttcctcctc tcctgactac tcccagtcat agctgtccct cttctcttat ggagatccct
     4021 cgacctgcag cccaagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
     4081 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
     4141 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     4201 gaaacctgtc gtgccagcgg atcgatccgc tgcattaatg aatcggccaa cgcgcgggga
     4261 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     4321 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     4381 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     4441 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     4501 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     4561 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     4621 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     4681 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     4741 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     4801 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     4861 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     4921 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     4981 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     5041 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     5101 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     5161 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     5221 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     5281 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     5341 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     5401 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     5461 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     5521 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     5581 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     5641 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     5701 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     5761 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     5821 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     5881 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     5941 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     6001 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     6061 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     6121 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     6181 gggttccgcg cacatttccc cgaaaagtgc cacctg
//
