LOCUS       sequence-237764-        4077 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses human IFITM3-N21del 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 4077)
  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 4077)
  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: IFITM3-N21del-V93-TAG
FEATURES             Location/Qualifiers
     source          1..4077
                     /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"
     polyA_signal    886..1020
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(910..929)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     964..983
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1131..1147)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1131..1147)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1144..1166)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1155..1171
                     /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(1179..1185,1186..1203,1204..1209))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1224..1245
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1362..1379)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1533..2121)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1613..1632)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2292..3083,3084..3152))
                     /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     2915..2934
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3153..3257)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3325..3343
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3381..3403)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3503..3522
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3716..3738
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3730..3747
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3731..3747
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        3774..4077
                     /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 gaaaaaccag aaagttaact ggtaagttta gtctttttgt
      361 cttttatttc aggtcccgga tccggtggtg gtgcaaatca aagaactgct cctcagtgga
      421 tgttgccttt acttctaggc ctgtacggaa gtgttacttc tgctctaaaa gctgcggaat
      481 tgtacccgcg ggcccaccat ggagatgctc aaggaggagc acgaggtggc tgtgctgggg
      541 gcgccccaca accctgctcc cccgacgtcc accgtgatcc acatccgcag cgagacctcc
      601 gtgcccgacc atgtcgtctg gtccctgttc aacaccctct tcatgaaccc ctgctgcctg
      661 ggcttcatag cattcgccta ctccgtgaag tctagggaca ggaagatggt tggcgactag
      721 accggggccc aggcctatgc ctccaccgcc aagtgcctga acatctgggc cctgattctg
      781 ggcatcctca tgaccattct gctcatcgtc atcccagtgc tgatcttcca ggcctatgga
      841 taataataag tcgaccgaga tctctcgagg taccgcggcc gcggggatcc agacatgata
      901 agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt
      961 tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt
     1021 aacaacaaca attgcattca ttttatgttt caggttcagg gggaggtgtg ggaggttttt
     1081 tcggatcctc tagagtcgat ctgcaggcat gctagcttgg cgtaatcatg gtcatagctg
     1141 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
     1201 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
     1261 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     1321 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     1381 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     1441 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     1501 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     1561 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     1621 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     1681 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     1741 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     1801 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     1861 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     1921 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     1981 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     2041 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     2101 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     2161 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     2221 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     2281 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     2341 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     2401 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     2461 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     2521 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     2581 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     2641 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgtta
     2701 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     2761 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     2821 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     2881 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     2941 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     3001 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     3061 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     3121 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     3181 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     3241 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt
     3301 attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt
     3361 ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt cacagcttgt
     3421 ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg
     3481 tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccatatg
     3541 cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg ccattcgcca
     3601 ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag
     3661 ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg gttttcccag
     3721 tcacgacgtt gtaaaacgac ggccagtgag ttcgagcttg catgcctgca ggtcgttaca
     3781 taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca
     3841 ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg
     3901 gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg
     3961 ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc
     4021 ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatg
//
