LOCUS       sequence-458388-        9002 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of S protein of SARS-CoV2 under CMV promoter, 
            tagged with 6His and released extracellular environment after 
            expression for protein purification.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9002)
  AUTHORS   Ozcelik CE, Araz CZ, Yilmaz O, Gulyuz S, Ozdamar P, Salmanli E, 
            Ozkul A, Seker UOS
  TITLE     Screening Peptide Drug Candidates To Neutralize Whole Viral Agents: 
            A Case Study with Severe Acute Respiratory Syndrome Coronavirus 2 
            (SARS-CoV-2).
  JOURNAL   ACS Pharmacol Transl Sci. 2024 Mar 21;7(4):1032-1042. doi: 
            10.1021/acsptsci.3c00317. eCollection 2024 Apr 12.
  PUBMED    38633598
REFERENCE   2  (bases 1 to 9002)
  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/acsptsci.3c00317"; journalName: "ACS Pharmacol Transl Sci";
            date: "2024-03-21- 21"; volume: "7"; issue: "4"; pages: "1032-1042"
COMMENT     Sequence Label: AP2u2-CMV-S_protein
FEATURES             Location/Qualifiers
     source          1..9002
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        17..396
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        397..600
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     551..571
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     645..664
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        645..663
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             complement(2021..2038)
                     /codon_start=1
                     /product="tetracysteine peptide that binds biarsenical 
                     labeling reagents"
                     /label=tetracysteine tag
                     /translation="CCPGCC"
     CDS             4351..4368
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             4462..4479
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     intron          4560..4789
                     /label=chimeric intron
                     /note="chimera between introns from adenovirus and 
                     immunoglobulin heavy chain genes"
     primer_bind     4772..4791
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     misc_feature    join(4862..5436,5437..5439,5440..5448)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(5029..5046)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     5256..5275
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             5471..6274
                     /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="MGSAIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQ
                     GRPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQD
                     LLSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE
                     EHQGLAPAELFARLKARMPDGDDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQ
                     DIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5534..5553)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6144..6163
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6404..6525
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6441..6460)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(6627..6643)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6627..6643)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6640..6662)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6651..6667
                     /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(6675..6681,6682..6699,6700..6705))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6720..6741
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6858..6875)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7029..7617)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7109..7128)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7788..8579,8580..8648))
                     /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     8411..8430
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8649..8753)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(8828..8847)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gccagatata cgcgttgaca ttgattattg actagttatt aatagtaatc aattacgggg
       61 tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg
      121 cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata
      181 gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc
      241 cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac
      301 ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg
      361 cagtacatct acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc
      421 aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc
      481 aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc
      541 gccccattga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct
      601 ctctggctaa ctagagaacc cactgcttac tggcttatcg aaattaatac gactcactat
      661 agggagaccc aagcttggta ccgagctcgg atcgatatcg ccgccaccat gttcgtgttc
      721 ctggtgctgc tgcccctggt ctccagccag tgtgtgaacc tgacaacccg gacacagctg
      781 cctcctgcct acacaaacag cttcacccgt ggcgtctact accccgacaa ggtcttccgg
      841 agtagcgtgc tccacagcac acaggatctg ttcctgccct tcttcagcaa tgtcacctgg
      901 ttccacgcca tccacgtgag cggcaccaac ggcacaaaga gattcgacaa ccccgtgctg
      961 cctttcaacg acggcgtgta cttcgccagt actgagaaga gcaacatcat cagaggatgg
     1021 atcttcggta ccaccctgga ctccaaaacc cagagcctgc tgatcgtgaa taatgccaca
     1081 aacgtggtga tcaaagtgtg tgaattccag ttttgtaatg atccatttct cggcgtatac
     1141 taccacaaga acaacaaaag ctggatggaa tccgagttca gagtgtatag ctctgctaac
     1201 aactgcacct tcgagtacgt gtcccagcct ttcctgatgg acctggaagg caagcagggc
     1261 aacttcaaga acctgagaga atttgtgttc aaaaacatcg atggctattt caagatctac
     1321 agcaagcaca cccctatcaa cctggtgcgg gatcttcccc agggcttcag cgccctggag
     1381 cctctggtgg acctgccgat cggaatcaac atcaccagat tccagaccct gctggccctg
     1441 cacagatctt acctgacccc tggcgacagc agcagtggct ggacagctgg agccgccgct
     1501 tattacgtgg gctacctgca gcctagaacc ttcctgctga aatacaacga gaacggaacc
     1561 atcaccgacg ctgttgattg cgctcttgat cccctgtctg aaaccaaatg caccctgaag
     1621 agctttaccg tggaaaaggg catatatcag acatccaact tcagagtgca gcctacagag
     1681 agcatcgtgc ggttccctaa tatcacgaac ctgtgtcctt tcggcgaggt gttcaacgcc
     1741 acccggttcg ccagcgtgta cgcctggaac cggaagagga tcagcaattg cgtggccgat
     1801 tacagcgtgc tgtacaacag cgccagcttc agcactttca agtgctacgg cgtgtctcct
     1861 accaagctga acgacctgtg cttcaccaac gtgtacgccg actcttttgt gatcagaggc
     1921 gacgaggtgc ggcagattgc ccctggccag acaggaaaga tcgcggatta caactacaag
     1981 ctgcctgatg acttcaccgg ctgcgtgatc gcctggaata gcaacaacct ggacagcaag
     2041 gtgggaggaa actacaacta cctgtaccgg ctgtttagaa agtctaacct gaagcctttc
     2101 gagcgggaca tcagcaccga gatataccag gccggctcaa caccttgcaa cggcgtggaa
     2161 ggattcaatt gttactttcc cctccaatct tacggcttcc agccaacaaa tggcgtcggc
     2221 taccagccgt acagagtcgt tgtgctctct ttcgagctgc tgcatgcccc tgccaccgtg
     2281 tgcggcccca agaaatccac caatctggtg aagaacaagt gcgtgaactt taactttaac
     2341 ggacttaccg gcacaggcgt tctgacggaa agcaacaaga agtttctgcc attccaacag
     2401 ttcggcagag acatcgccga caccacagat gccgtgaggg accctcaaac cctggaaata
     2461 ctcgacatca caccctgtag cttcggcgga gtgagcgtga tcactcccgg caccaacacc
     2521 tccaaccaag tggcagtgct gtaccaggac gtgaactgca ccgaggtccc cgtggccatc
     2581 cacgccgacc agctgacccc tacctggaga gtttactcca cagggagcaa cgttttccag
     2641 acccgggccg gctgtctgat cggcgccgag cacgtgaaca attcctacga gtgtgacatc
     2701 cccatcggcg ccggcatttg cgcaagctac cagacgcaga caaacagccc tgcttctgtg
     2761 gctagccaat ctatcatcgc ctacaccatg tctctcggcg ccgagaacag cgtggcttat
     2821 agcaacaaca gcatcgccat cccaaccaac ttcacaatca gcgtgacaac cgaaatcctg
     2881 cccgtgagca tgaccaagac ctccgtggat tgcacaatgt acatctgcgg cgacagcaca
     2941 gaatgctcta acctgctgct gcagtacggc agcttttgca cccaactgaa cagagccctg
     3001 accggcatcg ccgtggaaca ggataagaac acacaggagg tgttcgccca ggtgaaacag
     3061 atctacaaga cacctcccat caaagacttc ggcggattta acttctctca gatcctgcca
     3121 gaccccagca aacctagcaa gcggagcttc atcgaggacc tgctgttcaa caaggtgacc
     3181 ctggccgacg ctggatttat caagcagtac ggcgattgcc tgggcgacat cgctgccaga
     3241 gatctgatct gcgcccagaa gttcaatggc ctgacagtgc ttccgcctct gctgacagac
     3301 gagatgatcg cccagtacac cagcgctctg cttgccggca ccatcaccag cggatggacc
     3361 tttggagccg gcgccgccct gcaaatcccc ttcgccatgc agatggccta cagattcaac
     3421 ggaatcggcg tgacccagaa cgtgctgtac gaaaaccaga agctgattgc caatcagttc
     3481 aactccgcca tcggaaaaat ccaagactct ctgagctcca ccgccagcgc cctgggaaag
     3541 ctgcaagacg tggtcaacca gaacgcccag gccctgaaca ccctggttaa gcagctgagc
     3601 tctaacttcg gcgccatcag ctcagtgctg aatgacatcc tcagcagact ggaccctccc
     3661 gaggccgagg tccagatcga tagactcatc acaggtagac tgcagagcct gcaaacctac
     3721 gtcacacagc agctgatcag agccgccgag atcagagcca gcgccaacct ggctgccaca
     3781 aaaatgagcg aatgcgtgct gggccagagc aagcgcgtgg atttctgcgg caagggctac
     3841 cacctgatgt cttttccaca gagcgctccc cacggcgtgg tgttcctgca cgtgacctac
     3901 gtgcctgctc aagagaagaa ctttaccacc gcccctgcga tctgtcacga cggcaaggcc
     3961 catttccctc gggaaggcgt gttcgtgagc aatggtacac actggttcgt gacccaaaga
     4021 aacttctacg agcctcagat catcaccacc gacaacactt tcgtctccgg gaactgcgac
     4081 gtggtgatcg gcatcgtgaa caacaccgtg tacgaccctc tgcagccaga gctggacagc
     4141 tttaaggagg aactggacaa gtacttcaag aaccacacca gtcctgatgt ggacctgggt
     4201 gatattagcg gcatcaacgc cagcgtggtg aatatccaga aggaaatcga cagactgaac
     4261 gaggtggcca agaacctgaa cgaatccctg atcgacctgc aggagctggg taaatacgag
     4321 cagtacataa agtggcctgg aggcagcggt cttgtccctc ggggatcacc tgggtctggc
     4381 tacatacctg aagccccacg cgatggccaa gcctacgtaa ggaaagatgg tgaatgggtg
     4441 ttgctttcaa cctttctcgg ccaccaccac catcatcatt aataaggatc cgcggccgca
     4501 tagataactg atccagtgtg ctggaattaa ttcgctgtct gcgagggcca gctgttgggg
     4561 tgagtactcc ctctcaaaag cgggcatgac ttctgcgcta agattgtcag tttccaaaaa
     4621 cgaggaggat ttgatattca cctggcccgc ggtgatgcct ttgagggtgg ccgcgtccat
     4681 ctggtcagaa aagacaatct ttttgttgtc aagcttgagg tgtggcaggc ttgagatctg
     4741 gccatacact tgagtgacaa tgacatccac tttgcctttc tctccacagg tgtccactcc
     4801 caggtccaac tgcaggtcga gcatgcatct agggcggcca attccgcccc tctccccccc
     4861 ccccctctcc ctcccccccc cctaacgtta ctggccgaag ccgcttggaa taaggccggt
     4921 gtgcgtttgt ctatatgtta ttttccacca tattgccgtc ttttggcaat gtgagggccc
     4981 ggaaacctgg ccctgtcttc ttgacgagca ttcctagggg tctttcccct ctcgccaaag
     5041 gaatgcaagg tctgttgaat gtcgtgaagg aagcagttcc tctggaagct tcttgaagac
     5101 aaacaacgtc tgtagcgacc ctttgcaggc agcggaaccc cccacctggc gacaggtgcc
     5161 tctgcggcca aaagccacgt gtataagata cacctgcaaa ggcggcacaa ccccagtgcc
     5221 acgttgtgag ttggatagtt gtggaaagag tcaaatggct ctcctcaagc gtattcaaca
     5281 aggggctgaa ggatgcccag aaggtacccc attgtatggg atctgatctg gggcctcggt
     5341 gcacatgctt tacatgtgtt tagtcgaggt taaaaaaacg tctaggcccc ccgaaccacg
     5401 gggacgtggt tttcctttga aaaacacgat gataagcttg ccacaacccg ggataattcc
     5461 tgcagccaat atgggatcgg ccattgaaca agatggattg cacgcaggtt ctccggccgc
     5521 ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc
     5581 cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc
     5641 cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg
     5701 cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt
     5761 gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc
     5821 catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga
     5881 ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga
     5941 tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct
     6001 caaggcgcgc atgcccgacg gcgatgatct cgtcgtgacc catggcgatg cctgcttgcc
     6061 gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt
     6121 ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg
     6181 cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat
     6241 cgccttctat cgccttcttg acgagttctt ctgaggggat caattctcta gataactgat
     6301 cataatcagc cataccacat ttgtagaggt tttacttgct ttaaaaaacc tcccacacct
     6361 ccccctgaac ctgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc
     6421 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     6481 actgcattct agttgtggtt tgtccaaact catcaatgta tcttaacgcg tcgagtgcat
     6541 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc
     6601 tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg
     6661 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
     6721 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     6781 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     6841 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     6901 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     6961 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     7021 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     7081 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     7141 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     7201 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     7261 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     7321 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     7381 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     7441 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     7501 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     7561 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     7621 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     7681 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     7741 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     7801 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     7861 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     7921 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     7981 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     8041 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     8101 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     8161 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     8221 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     8281 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     8341 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     8401 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     8461 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     8521 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     8581 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     8641 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     8701 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     8761 ccccgaaaag tgccacctga cgtcgacgga tcgggagatc tcccgatccc ctatggtgca
     8821 ctctcagtac aatctgctct gatgccgcat agttaagcca gtatctgctc cctgcttgtg
     8881 tgttggaggt cgctgagtag tgcgcgagca aaatttaagc tacaacaagg caaggcttga
     8941 ccgacaattg catgaagaat ctgcttaggg ttaggcgttt tgcgctgctt cgcgatgtac
     9001 gg
//
