LOCUS       sequence-408102-        7387 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses H3_Hong Kong/1/1968 in the surface of  mammalian cells
             .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7387)
  TITLE     Human expression plasmids for Hemagglutinin surface expression
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7387)
  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            "Unpublished"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: HA Display H3_A/Hong Kong/1/1968
FEATURES             Location/Qualifiers
     source          1..7387
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    1007..1128
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1044..1063)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1098..1117
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1177..1193)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1177..1193)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1190..1212)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1201..1217
                     /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(1225..1231,1232..1249,1250..1255))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1270..1291
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1408..1425)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1579..2167)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1659..1678)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2338..3129,3130..3198))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTHLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     2961..2980
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3199..3302)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3378..3397)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        3569..3948
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3949..4152
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4103..4123
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    4154..4172
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4175..4193
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     4203..4227
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          4346..4478
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     4461..4480
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        4523..4541
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     sig_peptide     4587..4646
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     CDS             4989..5015
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="This is a variant of the standard HA tag, which has 
                     sequence YPYDVPDYA. Some anti-HA antibodies may not 
                     recognize this variant."
                     /translation="YPYDVPDYA"
     CDS             4989..5015
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             4995..5021
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="This is a variant of the standard HA tag, which has 
                     sequence YPYDVPDYA. Some anti-HA antibodies may not 
                     recognize this variant."
                     /translation="YDVPDYASL"
     CDS             6312..6377
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             6384..6407
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             6414..6437
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             6480..6626
                     /codon_start=1
                     /product="transmembrane domain from platelet derived growth
                     factor receptor beta"
                     /label=PDGFR-beta TM domain
                     /translation="AVGQDTQEVIVVPHSLPFKVVVISAILALVVLTIISLIILIMLWQ
                     KKPR"
     primer_bind     complement(6673..6690)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6679..6903
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(7036..7055)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    7187..7234
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     CDS             join(7242..7387,1..877)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             join(7242..7387,1..877)
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
ORIGIN
        1 aaatagctgc gccgatggtt tctacaaaga tcgttatgtt tatcggcact ttgcatcggc
       61 cgcgctcccg attccggaag tgcttgacat tggggaattc agcgagagcc tgacctattg
      121 catctcccgc cgtgcacagg gtgtcacgtt gcaagacctg cctgaaaccg aactgcccgc
      181 tgttctgcag ccggtcgcgg aggccatgga tgcgatcgct gcggccgatc ttagccagac
      241 gagcgggttc ggcccattcg gaccgcaagg aatcggtcaa tacactacat ggcgtgattt
      301 catatgcgcg attgctgatc cccatgtgta tcactggcaa actgtgatgg acgacaccgt
      361 cagtgcgtcc gtcgcgcagg ctctcgatga gctgatgctt tgggccgagg actgccccga
      421 agtccggcac ctcgtgcacg cggatttcgg ctccaacaat gtcctgacgg acaatggccg
      481 cataacagcg gtcattgact ggagcgaggc gatgttcggg gattcccaat acgaggtcgc
      541 caacatcttc ttctggaggc cgtggttggc ttgtatggag cagcagacgc gctacttcga
      601 gcggaggcat ccggagcttg caggatcgcc gcggctccgg gcgtatatgc tccgcattgg
      661 tcttgaccaa ctctatcaga gcttggttga cggcaatttc gatgatgcag cttgggcgca
      721 gggtcgatgc gacgcaatcg tccgatccgg agccgggact gtcgggcgta cacaaatcgc
      781 ccgcagaagc gcggccgtct ggaccgatgg ctgtgtagaa gtactcgccg atagtggaaa
      841 ccgacgcccc agcactcgtc cgagggcaaa ggaatagcac gtactacgag atttcgattc
      901 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat
      961 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc
     1021 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     1081 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat
     1141 accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     1201 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     1261 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     1321 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     1381 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     1441 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     1501 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     1561 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     1621 acgctcaagt cggaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     1681 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     1741 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     1801 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     1861 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     1921 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     1981 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     2041 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     2101 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     2161 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     2221 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     2281 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     2341 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     2401 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     2461 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     2521 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     2581 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     2641 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     2701 ctccggttcc caacgatcaa ggtgagttac atgatccccc atgttgtgca aaaaagcggt
     2761 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     2821 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     2881 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     2941 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     3001 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     3061 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     3121 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     3181 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     3241 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     3301 gcacacattt ccccgaaaag tgccacctga cgtcgacgga tcgggagatc tcccgatccc
     3361 ctatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca gtatctgctc
     3421 cctgcttgtg tgttggaggt cgctgagtag tgcgcgagca aaatttaagc tacaacaagg
     3481 caaggcttga ccgacaattg catgaagaat ctgcttaggg ttaggcgttt tgcgctgctt
     3541 cgcgatgtac gggccagata tacgcgttga cattgattat tgactagtta ttaatagtaa
     3601 tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg
     3661 gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg
     3721 tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta
     3781 cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt
     3841 gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac
     3901 tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt
     3961 tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac
     4021 cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt
     4081 cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat
     4141 ataagcagag ctctccctat cagtgataga gatctcccta tcagtgatag agatcgtcga
     4201 cgagctcgtt tagtgaaccg tcagatcgcc tggagacgcc atccacgctg ttttgacctc
     4261 catagaagac accgggaccg atccagcctc cggactctag cgtttaaact taagctgcag
     4321 aagttggtcg tgaggcactg ggcaggtaag tatcaaggtt acaagacagg tttaaggaga
     4381 ccaatagaaa ctgggcttgt cgagacagag aagactcttg cgtttctgat aggcacctat
     4441 tggtcttact gacatccact ttgcctttct ctccacaggt gtccactccc agttcaatta
     4501 cagctcttaa ggctagagta cttaatacga ctcactatag gctagcctcg agaattcacg
     4561 cgtggtacct ctagagtcga cccgggatgg agacagacac actcctgcta tgggtactgc
     4621 tgctctgggt tccaggttcc actggtgaca tgaagaccat catcgccctg agctacatct
     4681 tctgcctggc cctgggccag gacctgcccg gcaacgacaa cagcaccgcc accctgtgcc
     4741 tgggccacca cgccgtgccc aacggcaccc tggtgaagac catcaccgac gaccagatcg
     4801 aggtgaccaa cgccaccgag ctggtgcaga gcagcagcac cggcaagatc tgcaacaacc
     4861 cccaccggat cctggacggc atcgactgca ccctgatcga cgccctgctg ggcgaccccc
     4921 actgcgacgt gttccagaac gagacctggg acctgttcgt ggagcggagc aaggccttca
     4981 gcaactgcta cccctacgac gtgcccgact acgccagcct gcggagcctg gtggccagca
     5041 gcggcaccct ggagttcatc accgagggct tcacctggac cggcgtgacc cagaacggcg
     5101 gcagcaacgc ctgcaagcgg ggccccggca gcggcttctt cagccggctg aactggctga
     5161 ccaagagcgg cagcacctac cccgtgctga acgtgaccat gcccaacaac gacaacttcg
     5221 acaagctgta catctggggc gtgcaccacc ccagcaccaa ccaggagcag accagcctgt
     5281 acgtgcaggc cagcggccgg gtgaccgtga gcacccggcg gagccagcag accatcatcc
     5341 ccaacatcgg cagccggccc tgggtgcggg gcctgagcag ccggatcagc atctactgga
     5401 ccatcgtgaa gcccggcgac gtgctggtga tcaacagcaa cggcaacctg atcgcccccc
     5461 ggggctactt caagatgcgg accggcaaga gcagcatcat gcggagcgac gcccccatcg
     5521 atacatgtat cagcgagtgc atcaccccca acggcagcat ccccaacgac aagcccttcc
     5581 agaacgtgaa caagatcacc tacggcgcct gccccaagta cgtgaagcag aacaccctga
     5641 agctggccac cggcatgcgg aacgtgcccg agaagcagac ccggggcctg ttcggcgcca
     5701 tcgccggctt catcgagaac ggctgggagg gcatgatcga cggctggtac ggcttccggc
     5761 accagaacag cgagggcacc ggccaggccg ccgacctgaa gagcacccag gccggcatcg
     5821 accagatcaa cggcaagctg aaccgggtga tcgagaagac caacgagaag ttccaccaga
     5881 tcgagaagga gttcagcgag gtggagggcc ggatccagga cctggagaag tacgtggagg
     5941 acaccaagat cgacctgtgg agctacaacg ccgagctgct ggtggccctg gagaaccagc
     6001 acaccatcga cctgaccgac agcgagatga acaagctgtt cgagaagacc cggcggcagc
     6061 tgcgggagaa cgccgaggac atgggcaacg gctgcttcaa gatctaccac aagtgcgaca
     6121 acgcctgcat cgagagcatc cggaacggca cctacgacca cgacgtgtac cgggacgagg
     6181 ccctgaacaa ccggttccag atcaagggcg tgcagggatc aggatacatc cccgaggccc
     6241 ccagagatgg ccaggcctac gtgcggaagg acggcgagtg ggtactgctg agcacattcc
     6301 tgggcagatc cgactacaaa gaccatgacg gtgattataa agatcatgac atcgattaca
     6361 aggatgacga tgacaagagc gcctggtccc acccccagtt cgagaaggga tccctggagg
     6421 tgctgttcca gggcccaggc ggcggcggta gtggaggggg cggatctggc ggctcagctg
     6481 ctgtgggcca ggacacgcag gaggtcatcg tggtgccaca ctccttgccc tttaaggtgg
     6541 tggtgatctc agccatcctg gccctggtgg tgctcaccat catctccctt atcatcctca
     6601 tcatgctttg gcagaagaag ccacgttaac ggccgctcga gtctagaggg cccgtttaaa
     6661 cccgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc
     6721 ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg
     6781 aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg
     6841 acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta
     6901 tggcttctga ggcggaaaga accagctggg gctctagggg gtatccccac gcgccctgta
     6961 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     7021 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     7081 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     7141 acctcgaccc caaaaaactt gattagggtg atggttcacg tacctagaag ttcctattcc
     7201 gaagttccta ttctctagaa agtataggaa cttccttggc caaaaagcct gaactcaccg
     7261 cgacgtctgt cgagaagttt ctgatcgaaa agttcgacag cgtctccgac ctgatgcagc
     7321 tctcggaggg cgaagaatct cgtgctttca gcttcgatgt aggagggcgt ggatatgtcc
     7381 tgcgggt
//
