LOCUS       sequence-387128-        4184 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  N-terminal 3xHA mammalian expression plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4184)
  TITLE     tag plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4184)
  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            "Unpublished"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: pCS2+N-3xHA Cloning Vector
FEATURES             Location/Qualifiers
     source          1..4184
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        35..53
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     35..52
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             84..110
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             114..140
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             144..170
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    236..370
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(260..279)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     314..333
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(483..503)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(483..501)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(522..538)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(522..538)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(535..557)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    546..562
                     /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(570..576,577..594,595..600))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    615..636
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(753..770)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(924..1512)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1004..1023)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1683..2474,2475..2543))
                     /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     2306..2325
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2544..2648)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2674..3129)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2811..2832)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3023..3042
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3200..4184
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
ORIGIN
        1 cgccattctg cctggggacg tcggagcaag cttgatttag gtgacactat agacttgttc
       61 tttttgcagg atctaccatg ggatacccat acgatgttcc tgactatgcg ggctatccct
      121 atgacgtccc ggactatgca ggctatccat atgacgttcc agattacgct ggatcccatc
      181 gattcgaatt caaggcctct cgagcctcta gaactatagt gagtcgtatt acgtagatcc
      241 agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa
      301 atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa
      361 taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg gggaggtgtg
      421 ggaggttttt taattcgcgg ccgcggcgcc aatgcattgg gcccggtacc cagcttttgt
      481 tccctttagt gagggttaat tgcgcgcttg gcgtaatcat ggtcatagct gtttcctgtg
      541 tgaaattgtt atccgctcac aattccacac aacatacgag ccgggagcat aaagtgtaaa
      601 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
      661 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
      721 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
      781 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
      841 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
      901 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
      961 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     1021 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     1081 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     1141 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     1201 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     1261 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     1321 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     1381 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     1441 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     1501 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     1561 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     1621 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     1681 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     1741 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     1801 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     1861 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     1921 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     1981 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     2041 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     2101 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     2161 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     2221 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     2281 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     2341 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     2401 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     2461 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     2521 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     2581 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     2641 gttccgcgca catttccccg aaaagtgcca cctaaattgt aagcgttaat attttgttaa
     2701 aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca
     2761 aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga
     2821 acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc
     2881 agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc
     2941 gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc
     3001 cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg
     3061 caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac
     3121 agggcgcgtc ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg
     3181 cctcttcgct attacgccag tcgaccgcca attcaatatg gcgtatatgg actcatgcca
     3241 attcaatatg gtggatctgg acctgtgcca attcaatatg gcgtatatgg actcgtgcca
     3301 attcaatatg gtggatctgg accccagcca attcaatatg gcggacttgg caccatgcca
     3361 attcaatatg gcggacctgg cactgtgcca actggggagg ggtctacttg gcacggtgcc
     3421 aagtttgagg aggggtcttg gccctgtgcc aagtccgcca tattgaattg gcatggtgcc
     3481 aataatggcg gccatattgg ctatatgcca ggatcaatat ataggcaata tccaatatgg
     3541 ccctatgcca atatggctat tggccaggtt caatactatg tattggccct atgccatata
     3601 gtattccata tatgggtttt cctattgacg tagatagccc ctcccaatgg gcggtcccat
     3661 ataccatata tggggcttcc taataccgcc catagccact cccccattga cgtcaatggt
     3721 ctctatatat ggtctttcct attgacgtca tatgggcggt cctattgacg tatatggcgc
     3781 ctcccccatt gacgtcaatt acggtaaatg gcccgcctgg ctcaatgccc attgacgtca
     3841 ataggaccac ccaccattga cgtcaatggg atggctcatt gcccattcat atccgttctc
     3901 acgcccccta ttgacgtcaa tgacggtaaa tggcccactt ggcagtacat caatatctat
     3961 taatagtaac ttggcaagta cattactatt ggaagtacgc cagggtacat tggcagtact
     4021 cccattgacg tcaatggcgg taaatggccc gcgatggctg ccaagtacat ccccattgac
     4081 gtcaatgggg aggggcaatg acgcaaatgg gcgttccatt gacgtaaatg ggcggtaggc
     4141 gtgcctaatg ggaggtctat ataagcaatg ctcgtttagg gaac
//
