LOCUS       sequence-353670-        6745 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Expression vector for human IFITM3. Co-expresses BFP controlled by 
            an IRES element..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6745)
  AUTHORS   Prelli Bozzo C, Nchioua R, Volcic M, Koepke L, Kruger J, Schutz D, 
            Heller S, Sturzel CM, Kmiec D, Conzelmann C, Muller J, Zech F, Braun
            E, Gross R, Wettstein L, Weil T, Weiss J, Diofano F, Rodriguez 
            Alfonso AA, Wiese S, Sauter D, Munch J, Goffinet C, Catanese A, 
            Schon M, Boeckers TM, Stenger S, Sato K, Just S, Kleger A, Sparrer 
            KMJ, Kirchhoff F
  TITLE     IFITM proteins promote SARS-CoV-2 infection and are targets for 
            virus inhibition in vitro.
  JOURNAL   Nat Commun. 2021 Jul 28;12(1):4584.
  PUBMED    34321474
REFERENCE   2  (bases 1 to 6745)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Commun."; date: "2021-07-28"; volume: "12(1)"; pages: "4584"
COMMENT     Sequence Label: pCG_IFITM3-IRES_BFP
FEATURES             Location/Qualifiers
     source          1..6745
                     /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"
     5'UTR           598..637
                     /label=HSV TK 5'-UTR
                     /note="5' untranslated region from the herpes simplex virus
                     thymidine kinase gene"
     misc_feature    1114..1679
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    1130..1681
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1274..1291)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1501..1520
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(1684..1686,1687..1689,1690..2403)
                     /codon_start=1
                     /product="photostable monomeric blue variant of GFP (Ai et 
                     al., 2007)"
                     /label=mKalama1
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVEMDGDVNGRKFSVRGVGEGDATHGKLTL
                     KFICTSGKLPVPWPTLVTTLSYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GSYKTRAEVKFEGDTLVNRIVLKGTDFKEDGNILGHKLEYNMNVGNVYITADKQKNGIK
                     ANFEIRHNVEDGGVQLADHYQQNTPIGDGSVLLPDNHYLSVQVKLSKDPNEKRDHMVLL
                     EFRTAAGITPGMDELYK"
     intron          2444..3016
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     primer_bind     complement(3148..3167)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3213..3268
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3267..3286)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     rep_origin      3622..3757
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3863..3880)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4034..4622)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4114..4133)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4793..5584,5585..5653))
                     /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     5416..5435
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5654..5758)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5826..5844
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5882..5904)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6004..6023
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(6090..6545)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6227..6248)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6439..6458
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6671..6693
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6685..6702
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6686..6702
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6709..6728
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6709..6727
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
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 tccgggggat
      601 cttggtggcg tgaaactccc gcacctcttc ggccagcgcc ttgtagaagc gcgtaggcct
      661 tctagaccat gaatcacact gtccaaacct tcttctctcc tgtcaacagt ggccagcccc
      721 ccaactatga gatgctcaag gaggagcacg aggtggctgt gctgggggcg ccccacaacc
      781 ctgctccccc gacgtccacc gtgatccaca tccgcagcga gacctccgtg cccgaccatg
      841 tcgtctggtc cctgttcaac accctcttca tgaacccctg ctgcctgggc ttcatagcat
      901 tcgcctactc cgtgaagtct agggacagga agatggttgg cgacgtgacc ggggcccagg
      961 cctatgcctc caccgccaag tgcctgaaca tctgggccct gattctgggc atcctcatga
     1021 ccattctgct catcgtcatc ccagtgctga tcttccaggc ctatggatag acgcgtcgga
     1081 tcccggagta ctacaaagac tgctgacgcg aattccgccc ccccccctaa cgttactggc
     1141 cgaagccgct tggaataagg ccggtgtgcg tttgtctata tgttattttc caccatattg
     1201 ccgtcttttg gcaatgtgag ggcccggaaa cctggccctg tcttcttgac gagcattcct
     1261 aggggtcttt cccctctcgc caaaggaatg caaggtctgt tgaatgtcgt gaaggaagca
     1321 gttcctctgg aagcttcttg aagacaaaca acgtctgtag cgaccctttg caggcagcgg
     1381 aaccccccac ctggcgacag gtgcctctgc ggccaaaagc cacgtgtata agatacacct
     1441 gcaaaggcgg cacaacccca gtgccacgtt gtgagttgga tagttgtgga aagagtcaaa
     1501 tggctctcct caagcgtatt caacaagggg ctgaaggatg cccagaaggt accccattgt
     1561 atgggatctg atctggggcc tcggtgcaca tgctttacat gtgtttagtc gaggttaaaa
     1621 aacgtctagg ccccccgaac cacggggacg tggttttcct ttgaaaaaca cgatgataat
     1681 accatggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagatg
     1741 gacggcgacg taaacggccg caagttcagc gtgcgcggcg tgggcgaggg tgatgccacc
     1801 cacggcaagc tgaccctgaa gttcatctgc acctccggca agctgcccgt gccctggcct
     1861 accctcgtga ccaccctgtc ttacggcgtg cagtgcttct cccgctaccc cgaccacatg
     1921 aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc
     1981 ttcttcaagg acgacggcag ctacaagacc cgcgccgagg tgaagttcga gggcgacacc
     2041 ctggtgaacc gcatcgtgct gaagggcacc gacttcaagg aggacggtaa catcctgggg
     2101 cacaagctgg agtacaacat gaacgtcggg aacgtctata tcacggccga caagcagaag
     2161 aacggcatca aagcgaactt cgagatcagg cacaacgtcg aggacggcgg cgtgcagctc
     2221 gccgaccact accagcagaa cacccccatc ggcgacggct ccgtgctgct gcccgacaac
     2281 cactacctga gcgtgcaggt gaagctgagc aaagacccca acgagaagcg cgatcacatg
     2341 gtcctgctgg agttccgcac cgccgccggg atcactcccg gcatggacga gctgtacaag
     2401 taaggatcca ccggatcgag gtaaggatcc tgagaacttc agggtgagtt tggggaccct
     2461 tgattgttct ttctttttcg ctattgtaaa attcatgtta tatggagggg gcaaagtttt
     2521 cagggtgttg tttagaatgg gaagatgtcc cttgtatcac catggaccct catgataatt
     2581 ttgtttcttt cactttctac tctgttgaca accattgtct cctcttattt tcttttcatt
     2641 ttctgtaact ttttcgttaa actttagctt gcatttgtaa cgaattttta aattcacttt
     2701 tgtttatttg tcagattgta agtactttct ctaatcactt ttttttcaag gcaatcaggg
     2761 tatattatat tgtacttcag cacagtttta gagaacaatt gttataatta aatgataagg
     2821 tagaatattt ctgcatataa attctggctg gcgtggaaat attcttattg gtagaaacaa
     2881 ctacaccctg gtcatcatcc tgcctttctc tttatggtta caatgatata cactgtttga
     2941 gatgaggata aaatactctg agtccaaacc gggcccctct gctaaccatg ttcatgcctt
     3001 cttctctttc ctacagctcc tgggcaacgt gctggttgtt gtgctgtctc atcattttgg
     3061 caaagaattc actcctcagg tgcaggctgc ctatcagaag gtggtggctg gtgtggccaa
     3121 tgccctggct cacaaatacc actgagatct ttttccctct gccaaaaatt atggggacat
     3181 catgaagccc cttgagcatc tgacttctgg ctaataaagg aaatttattt tcattgcaat
     3241 agtgtgttgg aattttttgt gtctctcact cggaaggaca tatgggaggg caaatcattt
     3301 aaaacatcag aatgagtatt tggtttagag tttggcaaca tatgcccata tgctggctgc
     3361 catgaacaaa ggttggctat aaagaggtca tcagtatatg aaacagcccc ctgctgtcca
     3421 ttccttattc catagaaaag ccttgacttg aggttagatt ttttttatat tttgttttgt
     3481 gttatttttt tctttaacat ccctaaaatt ttccttacat gttttactag ccagattttt
     3541 cctcctctcc tgactactcc cagtcatagc tgtccctctt ctcttatgga gatccctcga
     3601 ggagcttttt gcaaaagcct aggcctccaa aaaagcctct tcactacttc tggaatagct
     3661 cagaggccga ggcggcctcg gcctctgcat aaataaaaaa aattagtcag ccatggggcg
     3721 gagaatgggc ggaactgggc ggagttaggg gcgggatggg cggagttagg ggcgggacta
     3781 tggttgctga ctaattgctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     3841 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     3901 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     3961 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     4021 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     4081 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     4141 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     4201 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     4261 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     4321 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     4381 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     4441 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     4501 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     4561 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     4621 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     4681 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     4741 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     4801 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     4861 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     4921 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     4981 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     5041 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     5101 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     5161 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     5221 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     5281 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     5341 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     5401 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     5461 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     5521 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     5581 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     5641 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     5701 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     5761 catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct
     5821 ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa
     5881 acctctgaca catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga
     5941 gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact
     6001 atgcggcatc agagcagatt gtactgagag tgcaccatat gcggtgtgaa ataccgcaca
     6061 gatgcgtaag gagaaaatac cgcatcagga aattgtaagc gttaatattt tgttaaaatt
     6121 cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat
     6181 cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa
     6241 gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg
     6301 cgatggccca ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa
     6361 agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc
     6421 gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag
     6481 tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg
     6541 cgcgtccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct
     6601 tcgctattac gccagctggc gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg
     6661 ccagggtttt cccagtcacg acgttgtaaa acgacggcca gtgaattgta atacgactca
     6721 ctatagggcg aattcgagct cgccc
//
