LOCUS       sequence-210496-        5941 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Fluorescent RILP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5941)
  AUTHORS   Mainou BA, Dermody TS
  TITLE     Transport to late endosomes is required for efficient reovirus 
            infection.
  JOURNAL   J Virol. 2012 Aug;86(16):8346-58. doi: 10.1128/JVI.00100-12. Epub 
            2012 Jun 6.
  PUBMED    22674975
REFERENCE   2  (bases 1 to 5941)
  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"; doi: 
            "10.1128/JVI.00100-12"; journalName: "J Virol"; date: "2012-08"; 
            volume: "86"; issue: "16"; pages: "8346-58"
COMMENT     Sequence Label: EGFP-RILP
FEATURES             Location/Qualifiers
     source          1..5941
                     /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"
     regulatory      547..556
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(553..555,556..558,559..1269)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(553..555,556..558,559..1269)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(598..619)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(859..878)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1206..1227
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2669..2790
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2706..2725)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2760..2779
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2797..3252)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2934..2955)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3146..3165
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3279..3383
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3385..3742
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3385..3576
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3408..3428)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3593..3728
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3777..4571
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             3777..4571
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3831..3850)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4441..4460
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4759..4778)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4803..4850
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5179..5767
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5668..5687
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
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 gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      541 ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
      601 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
      661 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
      721 ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc
      781 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
      841 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
      901 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
      961 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac
     1021 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc
     1081 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     1141 cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc
     1201 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     1261 ctgtacaagt ccggactcag atctcgagct caagcttcga attctgagcc caggagggcg
     1321 gcgcccgggg tgcctggctg ggggtctcgg gaggccgcgg ggtcggcatc ggccgcggag
     1381 cttgtgtacc atctagccgg ggccctgggc actgagctgc aggatctggc gcgccgtttc
     1441 gggccggagg cggcggccgg gctggtgccg ctagtggtgc gggcgctgga gctcttggaa
     1501 caggctgccg tggggcccgc cccggactcg ctgcaggtgt cggcgcagcc ggcggagcag
     1561 gagctgcggc ggctgcggga ggagaacgag cgcctccgca gggagctgcg cgcggggcca
     1621 caggaggagc gcgcgctgct gcggcagctc aaggaggtca cggaccgaca gcgggacgaa
     1681 ctccgggcgc acaaccgcga cctgcggcag cgcggccagg agaccgaggc gttgcaggag
     1741 cagctgcagc gcctcctgct ggtgaacgct gagctgcggc acaagctggc ggccatgcag
     1801 acccagctgc gcgccgcgca ggaccgcgag agggagcgcc agcagcctgg cgaagccgcg
     1861 actccgcagg ctaaagagcg agcgcggggg caggccgggc ggcccgggca ccagcacgga
     1921 caggagcccg aatgggcgac cgccggcgca ggcgccccag ggaaccctga ggacccggcg
     1981 gaggccgcgc agcagctcgg gcgcccctcg gaggcagggc agtgccgctt cagtcgggag
     2041 gagtttgagc agatccttca ggagcggaat gaactcaaag ccaaagtgtt cctgctcaag
     2101 gaggaactgg cctacttcca gcgggagctg ctcacagacc accgggtccc cggccttctg
     2161 ctcgaggcca tgaaggtggc tgtccggaag cagcggaaga agatcaaggc caagatgtta
     2221 gggacaccag aggaagcaga gagcagtgag gatgaggctg gcccatggat cctgctctcc
     2281 gatgacaagg gagaccatcc cccacccccg gagtccaaaa tacagagttt ctttggccta
     2341 tggtatcggg gtaaagctga atcctctgag gatgagacca gcagccctgc acccagcaag
     2401 ctagggggag aagaggaggc ccaaccacag tctccagctc ctgatccgcc ctgttctgcc
     2461 ctccacgaac acctttgtct gggggcctca gccgccccag aggcctgaga attctgcagt
     2521 cgacggtacc gcgggcccgg gatccaccgg atctagataa ctgatcataa tcagccatac
     2581 cacatttgta gaggttttac ttgctttaaa aaacctccca cacctccccc tgaacctgaa
     2641 acataaaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata atggttacaa
     2701 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     2761 tggtttgtcc aaactcatca atgtatctta acgcgtaaat tgtaagcgtt aatattttgt
     2821 taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg
     2881 gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt
     2941 ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct
     3001 atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt
     3061 gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa
     3121 agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc
     3181 tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc
     3241 tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     3301 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     3361 aataatattg aaaaaggaag agtcctgagg cggaaagaac cagctgtgga atgtgtgtca
     3421 gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     3481 caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     3541 aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc
     3601 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
     3661 tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt
     3721 tggaggccta ggcttttgca aagatcgatc aagagacagg atgaggatcg tttcgcatga
     3781 ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct
     3841 atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc
     3901 aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcaag
     3961 acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg
     4021 acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc
     4081 tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc
     4141 ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg
     4201 agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc
     4261 atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg cccgacggcg
     4321 aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc
     4381 gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag
     4441 cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg
     4501 tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg
     4561 agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc
     4621 atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt
     4681 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca
     4741 ccctaggggg aggctaactg aaacacggaa ggagacaata ccggaaggaa cccgcgctat
     4801 gacggcaata aaaagacaga ataaaacgca cggtgttggg tcgtttgttc ataaacgcgg
     4861 ggttcggtcc cagggctggc actctgtcga taccccaccg agaccccatt ggggccaata
     4921 cgcccgcgtt tcttcctttt ccccacccca ccccccaagt tcgggtgaag gcccagggct
     4981 cgcagccaac gtcggggcgg caggccctgc catagcctca ggttactcat atatacttta
     5041 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     5101 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     5161 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     5221 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     5281 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     5341 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     5401 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     5461 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     5521 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     5581 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     5641 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     5701 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     5761 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     5821 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgccatgca
     5881 ttagttatta atagtaatca attacggggt cattagttca tagcccatat atggagttcc
     5941 g
//
