LOCUS       sequence-328584-        5694 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes for first COVID protein - NSP1, NSP10, NSP11, and fragment 
            of NSP12 fused to nano-luciferase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5694)
  TITLE     NLuc CMV-Neo Flexi Vector pFC32K SARS-CoV2 5'Nsp1 PRF
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5694)
  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: 
            "Unpublished"
COMMENT     Sequence Label: NLuc CMV-Neo Flexi Vector pFC32K SARS-CoV2 5'N...
FEATURES             Location/Qualifiers
     source          1..5694
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        138..517
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        518..721
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     672..692
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     intron          857..989
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     972..991
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     primer_bind     1033..1052
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1033..1051
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2037..2549
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="VFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQRI
                     VLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGVT
                     PNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGWR
                     LCERILA"
     primer_bind     complement(2637..2655)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    2692..2813
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2703..2722)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2757..2776
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        3058..3415
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3058..3249
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3081..3101)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3266..3401
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        3430..3477
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3509..4303
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3563..3582)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4173..4192
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4367..4415
                     /note="synthetic polyadenylation signal"
     rep_origin      complement(4654..5242)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4734..4753)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    5358..5641
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to 
                     maintain plasmids as monomers"
ORIGIN
        1 tcaatattgg ccattagcca tattattcat tggttatata gcataaatca atattggcta
       61 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
      121 aatatgaccg ccatgttggc attgattatt gactagttat taatagtaat caattacggg
      181 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      241 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      301 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      361 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
      421 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
      481 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
      541 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      601 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaataaccc
      661 cgccccgttg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
      721 tggtttagtg aaccgtcaga tcactagaag ctttattgcg gtagtttatc acagttaaat
      781 tgctaacgca gtcagtgctt ctgacacaac agtctcgaac ttaagctgca gaagttggtc
      841 gtgaggcact gggcaggtaa gtatcaaggt tacaagacag gtttaaggag accaatagaa
      901 actgggcttg tcgagacaga gaagactctt gcgtttctga taggcaccta ttggtcttac
      961 tgacatccac tttgcctttc tctccacagg tgtccactcc cagttcaatt acagctctta
     1021 aggctagagt attaatacga ctcactatag ggctagcgat taaaggttta taccttccca
     1081 ggtaacaaac caaccaactt tcgatctctt gtagatctgt tctctaaacg aactttaaaa
     1141 tctgtgtggc tgtcactcgg ctgcatgctt agtgcactca cgcagtataa ttaataacta
     1201 attactgtcg ttgacaggac acgagtaact cgtctatctt ctgcaggctg cttacggttt
     1261 cgtccgtgtt gcagccgatc atcagcacat ctaggtttcg tccgggtgtg accgaaaggt
     1321 aagatggaga gccttgtccc tggtttcaac gagaaaacac acgtccaact cagtttgcct
     1381 gttttacagg ttcgcgacgt gctcgtacgt ggctttggag actccgtgga ggaggtctta
     1441 tcagaggcac gtcaacatct taaagatggc acttgtggct tagtagaagt tgaaaaaggc
     1501 gttttgcctc aacttgaaca gccctatgtg ttcatcaaac gttcggatgc tcgaactgca
     1561 cctcatggtc atgttatggt tgagctggta gcagaactcg aaggcattca gtacggtcgt
     1621 agtggtgaga cacttggtgt ccttgtccct catgtgggcg aaataccagt ggcttaccgc
     1681 aaggttcttc ttcgtaagaa cggtaataaa ggagctggtg gccatagtta cggcgccgat
     1741 ctaaagtcat ttgacttagg cgacgagctt ggcactgatc cttatgaaga ttttcaagaa
     1801 aactggaaca ctaaacatag cagtggtgtt acccgtgaac tcatgcgtga gcttaacgga
     1861 gggcccatgc ttcagtcagc tgatgcacaa tcgtttttaa acgggtttgc ggtgtaagtg
     1921 cagcccgtct tacaccgtgc ggcacaggca ctagtactga tgtcgtatac agggcttttg
     1981 acatctacaa tgataaagta gctggttttg ctaaattcgc tctcggctcg agcggcgtct
     2041 tcacactcga agatttcgtt ggggactggc gacagacagc cggctacaac ctggaccaag
     2101 tccttgaaca gggaggtgtg tccagtttgt ttcagaatct cggggtgtcc gtaactccga
     2161 tccaaaggat tgtcctgagc ggtgaaaatg ggctgaagat cgacatccat gtcatcatcc
     2221 cgtatgaagg tctgagcggc gaccaaatgg gccagatcga aaaaattttt aaggtggtgt
     2281 accctgtgga tgatcatcac tttaaggtga tcctgcacta tggcacactg gtaatcgacg
     2341 gggttacgcc gaacatgatc gactatttcg gacggccgta tgaaggcatc gccgtgttcg
     2401 acggcaaaaa gatcactgta acagggaccc tgtggaacgg caacaaaatt atcgacgagc
     2461 gcctgatcaa ccccgacggc tccctgctgt tccgagtaac catcaacgga gtgaccggct
     2521 ggcggctgtg cgaacgcatt ctggcgtaag gccgcgactc tagagtcgac ctgcaggcat
     2581 gcaagctgat ccggctgcta acaaagcccg aaaggaagct gagttggctg ctgccaccgc
     2641 tgagcaataa ctagcataac cccttggggc ggccgcttcg agcagacatg ataagataca
     2701 ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa
     2761 tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa gttaacaaca
     2821 acaattgcat tcattttatg tttcaggttc agggggagat gtgggaggtt tttttaagca
     2881 agtaaaacct ctacaaatgt ggtaaaatcg aattttaaca aaatattaac gcttacaatt
     2941 tcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atacgcggat
     3001 ctgcgcagca ccatggcctg aaataacctc tgaaagagga acttggttag gtaccttctg
     3061 aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc
     3121 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa
     3181 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     3241 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
     3301 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc
     3361 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct
     3421 taattaactg ttgacaatta atcatcggca tagtatatcg gcatagtata atacgacaag
     3481 gtgaggaact aaacccagga ggcagatcat gattgaacaa gatggattgc acgcaggttc
     3541 tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg
     3601 ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac
     3661 cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc
     3721 cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg
     3781 gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga
     3841 gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg
     3901 cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg
     3961 tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt
     4021 cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc
     4081 ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg
     4141 gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga
     4201 gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc
     4261 gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc
     4321 gaaatgaccg accaagcgac gcccaacctg ccatcacgat ggccgcaata aaatatcttt
     4381 attttcatta catctgtgtg ttggtttttt gtgtgaatcg atagcgataa ggatcctctt
     4441 tgcgcttgcg ttttcccttg tccagatagc ccagtagctg acattcatcc ggggtcagca
     4501 ccgtttctgc ggactggctt tctacccggt atcagctcac tcaaaggcgg taatacggtt
     4561 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     4621 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     4681 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     4741 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     4801 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     4861 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     4921 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     4981 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     5041 aggcagtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt
     5101 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     5161 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     5221 gcgcagaaaa aaaggatttc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     5281 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     5341 ctagatcctt ttatagtccg gaaatacagg aacgcacgct ggatggccct tcgctgggat
     5401 ggtgaaacca tgaaaaatgg cagcttcagt ggattaagtg ggggtaatgt ggcctgtacc
     5461 ctctggttgc ataggtattc atacggttaa aatttatcag gcgcgattgc ggcagttttt
     5521 cgggtggttt gttgccattt ttacctgtct gctgccgtga tcgcgctgaa cgcgttttag
     5581 cggtgcgtac aattaaggga ttatggtaaa tccacttact gtctgccctc gtagccatcg
     5641 agataaaccg cagtactccg gccacgatgc gtccggcgta gaggatcgag atct
//
