LOCUS       sequence-371-exp        6832 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6832)
  AUTHORS   Sherer NM, Lehmann MJ, Jimenez-Soto LF, Ingmundson A, Horner SM, 
            Cicchetti G, Allen PG, Pypaert M, Cunningham JM, Mothes W
  TITLE     Visualization of retroviral replication in living cells reveals 
            budding into multivesicular bodies.
  JOURNAL   Traffic 2003 Nov;4(11):785-801.
  PUBMED    14617360
REFERENCE   2  (bases 1 to 6832)
  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: "Traffic"; 
            date: "2003-11"; volume: "4(11)"; pages: "785-801"
COMMENT     Sequence Label: MLV Gag-RFP
FEATURES             Location/Qualifiers
     source          1..6832
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     misc_feature    609..665
                     /label=MCS
                     /note="multiple cloning site"
     intron          679..1154
                     /label=beta-globin intron
                     /note="internally truncated intron from human beta-globin"
     CDS             1212..1628
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1558..1580
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1596..1612
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     regulatory      2805..2814
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2811..3485
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     CDS             2811..3485
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /note="mammalian codon-optimized"
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     primer_bind     complement(2941..2959)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3209..3228
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     3403..3426
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     polyA_signal    3620..3741
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3657..3676)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3711..3730
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3748..4203)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3885..3906)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4097..4116
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4230..4334
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4336..4693
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4336..4527
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4359..4379)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4544..4679
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4728..5522
                     /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             4728..5522
                     /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(4782..4801)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5392..5411
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5710..5729)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5754..5801
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6130..6718
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6619..6638
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggactcag atctcgagct caagcttcga attctgcagt cgacggtacc gcgggcccgg
      661 gatcctgaga acttcagggt gagtctatgg gacccttgat gttttctttc cccttctttt
      721 ctatggttaa gttcatgtca taggaagggg agaagtaaca gggtacacat attgaccaaa
      781 tcagggtaat tttgcatttg taattttaaa aaatgctttc ttcttttaat atactttttt
      841 gtttatctta tttctaatac tttccctaat ctctttcttt cagggcaata atgatacaat
      901 gtatcatgcc tctttgcacc attctaaaga ataacagtga taatttctgg gttaaggcaa
      961 tagcaatatt tctgcatata aatatttctg catataaatt gtaactgatg taagaggttt
     1021 catattgcta atagcagcta caatccagct accattctgc ttttatttta tggttgggat
     1081 aaggctggat tattctgagt ccaagctagg cccttttgct aatcatgttc atacctctta
     1141 tcttcctccc acagctcctg ggcaacgtgc tggtctgtgt gctggcccat cactttggca
     1201 aagaattcat gggccagact gttaccactc ccttaagttt gaccttaggt cactggaaag
     1261 atgtcgagcg gatcgctcac aaccagtcgg tagatgtcaa gaagagacgt tgggttacct
     1321 tctgctctgc agaatggcca acctttaacg tcggatggcc gcgagacggc acctttaacc
     1381 gagacctcat cacccaggtt aagatcaagg tcttttcacc tggcccgcat ggacacccag
     1441 accaggtccc ctacatcgtg acctgggaag ccttggcttt tgacccccct ccctgggtca
     1501 agccctttgt acaccctaag cctccgcctc ctcttcctcc atccgccccg tctctccccc
     1561 ttgaacctcc tcgttcgacc ccgcctcgat cctcccttta tccagccctc actccttctc
     1621 taggcgccaa acctaaacct caagttcttt ctgacagtgg ggggccgctc atcgacctac
     1681 ttacagaaga ccccccgcct tatagggacc caagaccacc cccttccgac agggacggaa
     1741 atggtggaga agcgacccct gcgggagagg caccggaccc ctccccaatg gcatctcgcc
     1801 tacgtgggag acgggagccc cctgtggccg actccactac ctcgcaggca ttccccctcc
     1861 gcgcaggagg aaacggacag cttcaatact ggccgttctc ctcttctgac ctttacaact
     1921 ggaaaaataa taacccttct ttttctgaag atccaggtaa actgacagct ctgatcgagt
     1981 ctgttctcat cacccatcag cccacctggg acgactgtca gcagctgttg gggactctgc
     2041 tgaccggaga agaaaaacaa cgggtgctct tagaggctag aaaggcggtg cggggcgatg
     2101 atgggcgccc cactcaactg cccaatgaag tcgatgccgc ttttcccctc gagcgcccag
     2161 actgggatta caccacccag gcaggtagga accacctagt ccactatcgc cagttgctcc
     2221 tagcgggtct ccaaaacgcg ggcagaagcc ccaccaattt ggccaaggta aaaggaataa
     2281 cacaagggcc caatgagtct ccctcggcct tcctagagag acttaaggaa gcctatcgca
     2341 ggtacactcc ttatgaccct gaggacccag ggcaagaaac taatgtgtct atgtctttca
     2401 tttggcagtc tgccccagac attgggagaa agttagagag gttagaagat ttaaaaaaca
     2461 agacgcttgg agatttggtt agagaggcag aaaagatctt taataaacga gaaaccccgg
     2521 aagaaagaga ggaacgtatc aggagagaaa cagaggaaaa agaagaacgc cgtaggacag
     2581 aggatgagca gaaagagaaa gaaagagatc gtaggagaca tagagagatg agcaagctat
     2641 tggccactgt cgttagtgga cagaaacagg atagacaggg aggagaacga aggaggtccc
     2701 aactcgatcg cgaccagtgt gcctactgca aagaaaaggg gcactgggct aaagattgtc
     2761 ccaagaaacc acgaggacct cggggaccaa gaccgcaacc ggtcgccacc atggcctcct
     2821 ccgaggacgt catcaaggag ttcatgcgct tcaaggtgcg catggagggc tccgtgaacg
     2881 gccacgagtt cgagatcgag ggcgagggcg agggccgccc ctacgagggc acccagaccg
     2941 ccaagctgaa ggtgaccaag ggcggccccc tgcccttcgc ctgggacatc ctgtcccctc
     3001 agttccagta cggctccaag gcctacgtga agcaccccgc cgacatcccc gactacttga
     3061 agctgtcctt ccccgagggc ttcaagtggg agcgcgtgat gaacttcgag gacggcggcg
     3121 tggtgaccgt gacccaggac tcctccctgc aggacggcga gttcatctac aaggtgaagc
     3181 tgcgcggcac caacttcccc tccgacggcc ccgtaatgca gaagaagacc atgggctggg
     3241 aggcctccac cgagcggatg taccccgagg acggcgccct gaagggcgag atcaagatga
     3301 ggctgaagct gaaggacggc ggccactacg acgccgaggt caagaccacc tacatggcca
     3361 agaagcccgt gcagctgccc ggcgcctaca agaccgacat caagctggac atcacctccc
     3421 acaacgagga ctacaccatc gtggaacagt acgagcgcgc cgagggccgc cactccaccg
     3481 gcgccttgta caagtaaagc ggccgcgact ctagatcata atcagccata ccacatttgt
     3541 agaggtttta cttgctttaa aaaacctccc acacctcccc ctgaacctga aacataaaat
     3601 gaatgcaatt gttgttgtta acttgtttat tgcagcttat aatggttaca aataaagcaa
     3661 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     3721 caaactcatc aatgtatctt aaggcgtaaa ttgtaagcgt taatattttg ttaaaattcg
     3781 cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc
     3841 cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga
     3901 gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg
     3961 atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag
     4021 cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga
     4081 acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg
     4141 tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg
     4201 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     4261 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     4321 gaaaaaggaa gagtcctgag gcggaaagaa ccagctgtgg aatgtgtgtc agttagggtg
     4381 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     4441 agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     4501 tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc
     4561 gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc
     4621 cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct
     4681 aggcttttgc aaagatcgat caagagacag gatgaggatc gtttcgcatg attgaacaag
     4741 atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg
     4801 cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc
     4861 cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcaa gacgaggcag
     4921 cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca
     4981 ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat
     5041 ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata
     5101 cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac
     5161 gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc
     5221 tcgcgccagc cgaactgttc gccaggctca aggcgagcat gcccgacggc gaggatctcg
     5281 tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg
     5341 gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta
     5401 cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg
     5461 gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct
     5521 gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga
     5581 tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc
     5641 cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc accctagggg
     5701 gaggctaact gaaacacgga aggagacaat accggaagga acccgcgcta tgacggcaat
     5761 aaaaagacag aataaaacgc acggtgttgg gtcgtttgtt cataaacgcg gggttcggtc
     5821 ccagggctgg cactctgtcg ataccccacc gagaccccat tggggccaat acgcccgcgt
     5881 ttcttccttt tccccacccc accccccaag ttcgggtgaa ggcccagggc tcgcagccaa
     5941 cgtcggggcg gcaggccctg ccatagcctc aggttactca tatatacttt agattgattt
     6001 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     6061 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     6121 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     6181 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     6241 aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg
     6301 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     6361 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     6421 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     6481 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     6541 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     6601 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     6661 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     6721 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
     6781 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgccatgc at
//
