LOCUS       sequence-237722-        4191 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses human IFITM3-C71A 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-F8-TAG-C71A
FEATURES             Location/Qualifiers
     source          1..4191
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..204
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     155..175
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     201..225
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          342..438
                     /label=SV40 intron
                     /note="modified SV40 intron with splice donor and acceptor 
                     sites"
     CDS             502..528
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    1000..1134
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1024..1043)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1078..1097
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1245..1261)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1245..1261)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1258..1280)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1269..1285
                     /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(1293..1299,1300..1317,1318..1323))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1338..1359
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1476..1493)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1647..2235)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1727..1746)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2406..3197,3198..3266))
                     /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     3029..3048
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3267..3371)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3439..3457
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3495..3517)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3617..3636
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3830..3852
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3844..3861
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3845..3861
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        3888..4191
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
       61 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
      121 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
      181 tgggaggtct atataagcag agctcgttta gtgaaccgtc agatcgcctg gagacgccat
      241 ccacgctgtt ttgacctcca tagaagacac cgggaccgat ccagcctccg gactctagag
      301 gatccggtac tagaggaact gaaaaacaag aaagttaact ggtaagttta gtctttttgt
      361 cttttatttc aggtcccgga tccggtggtg gtgcaaatca aagaactgct cctcagtgga
      421 tgttgccttt acttctaggc ctgtacggaa gtgttacttc tgctctaaaa gctgcggaat
      481 tgtacccgcg ggcccaccat gtacccatac gatgttccag attacgctct tatggccatg
      541 gaggcccgaa ttcccatgaa tcacactgtc caaacctagt tctctcctgt caacagtggc
      601 cagcccccca actatgagat gctcaaggag gagcacgagg tggctgtgct gggggcgccc
      661 cacaaccctg ctcccccgac gtccaccgtg atccacatcc gcagcgagac ctccgtgccc
      721 gaccatgtcg tctggtccct gttcaacacc ctcttcatga accccgcctg cctgggcttc
      781 atagcattcg cctactccgt gaagtctagg gacaggaaga tggttggcga cgtgaccggg
      841 gcccaggcct atgcctccac cgccaagtgc ctgaacatct gggccctgat tctgggcatc
      901 ctcatgacca ttctgctcat cgtcatccca gtgctgatct tccaggccta tggataataa
      961 taagtcgacc gagatctctc gaggtaccgc ggccgcgggg atccagacat gataagatac
     1021 attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa
     1081 atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac
     1141 aacaattgca ttcattttat gtttcaggtt cagggggagg tgtgggaggt tttttcggat
     1201 cctctagagt cgatctgcag gcatgctagc ttggcgtaat catggtcata gctgtttcct
     1261 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
     1321 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
     1381 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     1441 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     1501 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     1561 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     1621 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     1681 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     1741 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     1801 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     1861 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     1921 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     1981 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     2041 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     2101 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     2161 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     2221 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     2281 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     2341 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     2401 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     2461 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     2521 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     2581 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     2641 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     2701 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     2761 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     2821 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     2881 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     2941 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     3001 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     3061 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     3121 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     3181 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     3241 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     3301 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     3361 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac cattattatc
     3421 atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt
     3481 gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc ttgtctgtaa
     3541 gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg
     3601 ggctggctta actatgcggc atcagagcag attgtactga gagtgcacca tatgcggtgt
     3661 gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggcgccattc gccattcagg
     3721 ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg ccagctggcg
     3781 aaagggggat gtgctgcaag gcgattaagt tgggtaacgc cagggttttc ccagtcacga
     3841 cgttgtaaaa cgacggccag tgagttcgag cttgcatgcc tgcaggtcgt tacataactt
     3901 acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg
     3961 acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat
     4021 ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct
     4081 attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg
     4141 gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat g
//
