LOCUS       sequence-322364-        5539 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes for first COVID protein - NSP1 fused to nano-luciferase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5539)
  TITLE     NLuc CMV-Neo Flexi Vector pFC32K SARS-CoV2 5'Nsp1 PRF
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5539)
  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            "Unpublished"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'Nsp1
FEATURES             Location/Qualifiers
     source          1..5539
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        66..445
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        446..649
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     600..620
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     intron          785..917
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     900..919
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     primer_bind     961..980
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        961..979
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1810..2322
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="VFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQRI
                     VLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGVT
                     PNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGWR
                     LCERILA"
     primer_bind     complement(2410..2428)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    2465..2586
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2476..2495)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2530..2549
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        2831..3188
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2831..3022
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2854..2874)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3039..3174
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        3203..3250
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3282..4076
                     /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(3336..3355)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3946..3965
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4140..4188
                     /note="synthetic polyadenylation signal"
     rep_origin      complement(4427..5015)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4507..4526)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    5131..5414
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to 
                     maintain plasmids as monomers"
ORIGIN
        1 tacgttgtat ctatatcata atatgtacat ttatattggc tcatgtccaa tatgaccgcc
       61 atgttggcat tgattattga ctagttatta atagtaatca attacggggt cattagttca
      121 tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc
      181 gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat
      241 agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt
      301 acatcaagtg tatcatatgc caagtccgcc ccctattgac gtcaatgacg gtaaatggcc
      361 cgcctggcat tatgcccagt acatgacctt acgggacttt cctacttggc agtacatcta
      421 cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacacca atgggcgtgg
      481 atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt
      541 gttttggcac caaaatcaac gggactttcc aaaatgtcgt aataaccccg ccccgttgac
      601 gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctg gtttagtgaa
      661 ccgtcagatc actagaagct ttattgcggt agtttatcac agttaaattg ctaacgcagt
      721 cagtgcttct gacacaacag tctcgaactt aagctgcaga agttggtcgt gaggcactgg
      781 gcaggtaagt atcaaggtta caagacaggt ttaaggagac caatagaaac tgggcttgtc
      841 gagacagaga agactcttgc gtttctgata ggcacctatt ggtcttactg acatccactt
      901 tgcctttctc tccacaggtg tccactccca gttcaattac agctcttaag gctagagtat
      961 taatacgact cactataggg ctagcgatta aaggtttata ccttcccagg taacaaacca
     1021 accaactttc gatctcttgt agatctgttc tctaaacgaa ctttaaaatc tgtgtggctg
     1081 tcactcggct gcatgcttag tgcactcacg cagtataatt aataactaat tactgtcgtt
     1141 gacaggacac gagtaactcg tctatcttct gcaggctgct tacggtttcg tccgtgttgc
     1201 agccgatcat cagcacatct aggtttcgtc cgggtgtgac cgaaaggtaa gatggagagc
     1261 cttgtccctg gtttcaacga gaaaacacac gtccaactca gtttgcctgt tttacaggtt
     1321 cgcgacgtgc tcgtacgtgg ctttggagac tccgtggagg aggtcttatc agaggcacgt
     1381 caacatctta aagatggcac ttgtggctta gtagaagttg aaaaaggcgt tttgcctcaa
     1441 cttgaacagc cctatgtgtt catcaaacgt tcggatgctc gaactgcacc tcatggtcat
     1501 gttatggttg agctggtagc agaactcgaa ggcattcagt acggtcgtag tggtgagaca
     1561 cttggtgtcc ttgtccctca tgtgggcgaa ataccagtgg cttaccgcaa ggttcttctt
     1621 cgtaagaacg gtaataaagg agctggtggc catagttacg gcgccgatct aaagtcattt
     1681 gacttaggcg acgagcttgg cactgatcct tatgaagatt ttcaagaaaa ctggaacact
     1741 aaacatagca gtggtgttac ccgtgaactc atgcgtgagc ttaacggagg ggctctcggc
     1801 tcgagcggcg tcttcacact cgaagatttc gttggggact ggcgacagac agccggctac
     1861 aacctggacc aagtccttga acagggaggt gtgtccagtt tgtttcagaa tctcggggtg
     1921 tccgtaactc cgatccaaag gattgtcctg agcggtgaaa atgggctgaa gatcgacatc
     1981 catgtcatca tcccgtatga aggtctgagc ggcgaccaaa tgggccagat cgaaaaaatt
     2041 tttaaggtgg tgtaccctgt ggatgatcat cactttaagg tgatcctgca ctatggcaca
     2101 ctggtaatcg acggggttac gccgaacatg atcgactatt tcggacggcc gtatgaaggc
     2161 atcgccgtgt tcgacggcaa aaagatcact gtaacaggga ccctgtggaa cggcaacaaa
     2221 attatcgacg agcgcctgat caaccccgac ggctccctgc tgttccgagt aaccatcaac
     2281 ggagtgaccg gctggcggct gtgcgaacgc attctggcgt aaggccgcga ctctagagtc
     2341 gacctgcagg catgcaagct gatccggctg ctaacaaagc ccgaaaggaa gctgagttgg
     2401 ctgctgccac cgctgagcaa taactagcat aaccccttgg ggcggccgct tcgagcagac
     2461 atgataagat acattgatga gtttggacaa accacaacta gaatgcagtg aaaaaaatgc
     2521 tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag ctgcaataaa
     2581 caagttaaca acaacaattg cattcatttt atgtttcagg ttcaggggga gatgtgggag
     2641 gtttttttaa gcaagtaaaa cctctacaaa tgtggtaaaa tcgaatttta acaaaatatt
     2701 aacgcttaca atttcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac
     2761 cgcatacgcg gatctgcgca gcaccatggc ctgaaataac ctctgaaaga ggaacttggt
     2821 taggtacctt ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa
     2881 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     2941 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     3001 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     3061 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     3121 ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     3181 tttgcaaaaa gcttaattaa ctgttgacaa ttaatcatcg gcatagtata tcggcatagt
     3241 ataatacgac aaggtgagga actaaaccca ggaggcagat catgattgaa caagatggat
     3301 tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac
     3361 agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc
     3421 tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc
     3481 tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag
     3541 cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc
     3601 ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg
     3661 atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc
     3721 ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc
     3781 cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga
     3841 cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca
     3901 tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg
     3961 atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg
     4021 ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg
     4081 gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gatggccgca
     4141 ataaaatatc tttattttca ttacatctgt gtgttggttt tttgtgtgaa tcgatagcga
     4201 taaggatcct ctttgcgctt gcgttttccc ttgtccagat agcccagtag ctgacattca
     4261 tccggggtca gcaccgtttc tgcggactgg ctttctaccc ggtatcagct cactcaaagg
     4321 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     4381 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     4441 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     4501 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     4561 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     4621 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     4681 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     4741 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     4801 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     4861 ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     4921 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     4981 agcagcagat tacgcgcaga aaaaaaggat ttcaagaaga tcctttgatc ttttctacgg
     5041 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     5101 aaaggatctt cacctagatc cttttatagt ccggaaatac aggaacgcac gctggatggc
     5161 ccttcgctgg gatggtgaaa ccatgaaaaa tggcagcttc agtggattaa gtgggggtaa
     5221 tgtggcctgt accctctggt tgcataggta ttcatacggt taaaatttat caggcgcgat
     5281 tgcggcagtt tttcgggtgg tttgttgcca tttttacctg tctgctgccg tgatcgcgct
     5341 gaacgcgttt tagcggtgcg tacaattaag ggattatggt aaatccactt actgtctgcc
     5401 ctcgtagcca tcgagataaa ccgcagtact ccggccacga tgcgtccggc gtagaggatc
     5461 gagatcttca atattggcca ttagccatat tattcattgg ttatatagca taaatcaata
     5521 ttggctattg gccattgca
//
