LOCUS       sequence-237724-        4191 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses human IFITM3 with unnatural amino acid incorporated site 
            specifically in response to the amber codon TAG in mammalian cells 
            when co-transfected with tRNA synthetase/tRNA pair..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4191)
  AUTHORS   Spence JS, He R, Hoffmann HH, Das T, Thinon E, Rice CM, Peng T, 
            Chandran K, Hang HC
  TITLE     IFITM3 directly engages and shuttles incoming virus particles to 
            lysosomes.
  JOURNAL   Nat Chem Biol. 2019 Jan 14. pii: 10.1038/s41589-018-0213-2. doi: 
            10.1038/s41589-018-0213-2.
  PUBMED    30643282
REFERENCE   2  (bases 1 to 4191)
  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: "Nat Chem 
            Biol. 2019 Jan 14. pii: 10.1038/s41589-018-0213-2. doi: 
            10.1038/s41589-018-0213-2."
COMMENT     Sequence Label: HA-IFITM3-V93-TAG
FEATURES             Location/Qualifiers
     source          1..4191
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     505..529
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          646..742
                     /label=SV40 intron
                     /note="modified SV40 intron with splice donor and acceptor 
                     sites"
     CDS             806..832
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    1304..1438
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1328..1347)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1382..1401
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1549..1565)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1549..1565)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1562..1584)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1573..1589
                     /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(1597..1603,1604..1621,1622..1627))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1642..1663
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1780..1797)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1951..2539)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2031..2050)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2710..3501,3502..3570))
                     /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     3333..3352
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3571..3675)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3743..3761
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3799..3821)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3921..3940
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4134..4156
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4148..4165
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4149..4165
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctcg tttagtgaac cgtcagatcg cctggagacg
      541 ccatccacgc tgttttgacc tccatagaag acaccgggac cgatccagcc tccggactct
      601 agaggatccg gtactagagg aactgaaaaa ccagaaagtt aactggtaag tttagtcttt
      661 ttgtctttta tttcaggtcc cggatccggt ggtggtgcaa atcaaagaac tgctcctcag
      721 tggatgttgc ctttacttct aggcctgtac ggaagtgtta cttctgctct aaaagctgcg
      781 gaattgtacc cgcgggccca ccatgtaccc atacgatgtt ccagattacg ctcttatggc
      841 catggaggcc cgaattccca tgaatcacac tgtccaaacc ttcttctctc ctgtcaacag
      901 tggccagccc cccaactatg agatgctcaa ggaggagcac gaggtggctg tgctgggggc
      961 gccccacaac cctgctcccc cgacgtccac cgtgatccac atccgcagcg agacctccgt
     1021 gcccgaccat gtcgtctggt ccctgttcaa caccctcttc atgaacccct gctgcctggg
     1081 cttcatagca ttcgcctact ccgtgaagtc tagggacagg aagatggttg gcgactagac
     1141 cggggcccag gcctatgcct ccaccgccaa gtgcctgaac atctgggccc tgattctggg
     1201 catcctcatg accattctgc tcatcgtcat cccagtgctg atcttccagg cctatggata
     1261 ataataagtc gaccgagatc tctcgaggta ccgcggccgc ggggatccag acatgataag
     1321 atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg
     1381 tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata aacaagttaa
     1441 caacaacaat tgcattcatt ttatgtttca ggttcagggg gaggtgtggg aggttttttc
     1501 ggatcctcta gagtcgatct gcaggcatgc tagcttggcg taatcatggt catagctgtt
     1561 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     1621 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     1681 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     1741 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     1801 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     1861 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     1921 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     1981 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     2041 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     2101 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     2161 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     2221 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     2281 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     2341 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     2401 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     2461 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     2521 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     2581 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     2641 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     2701 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     2761 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     2821 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     2881 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     2941 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     3001 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     3061 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     3121 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     3181 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     3241 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     3301 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     3361 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     3421 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     3481 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     3541 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     3601 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     3661 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat
     3721 tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt
     3781 cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct
     3841 gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg
     3901 tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc accatatgcg
     3961 gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc attcgccatt
     4021 caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct
     4081 ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt tttcccagtc
     4141 acgacgttgt aaaacgacgg ccagtgagtt cgagcttgca tgcctgcagg t
//
