LOCUS       sequence-148006-        5838 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 5838)
  AUTHORS   Lee HH, Elia N, Ghirlando R, Lippincott-Schwartz J, Hurley JH
  TITLE     Midbody targeting of the ESCRT machinery by a noncanonical coiled 
            coil in CEP55.
  JOURNAL   Science. 2008 Oct 24. 322(5901):576-80.
  PUBMED    18948538
REFERENCE   2  (bases 1 to 5838)
  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: "Science. 
            2008 Oct 24. 322(5901):576-80."
COMMENT     Sequence Label: mCherry-Tsg101
FEATURES             Location/Qualifiers
     source          1..5838
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        68..371
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        372..575
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     526..546
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      614..623
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             620..1294
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     CDS             620..1294
                     /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(750..768)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1018..1037
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     1212..1235
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     polyA_signal    2633..2754
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2670..2689)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2724..2743
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2761..3216)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2898..2919)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3110..3129
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3243..3347
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3349..3706
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3349..3540
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3372..3392)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3557..3692
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3741..4535
                     /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             3741..4535
                     /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(3795..3814)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4405..4424
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4723..4742)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4767..4814
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5143..5731
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5632..5651
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 catgcattag ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg
       61 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
      121 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt
      181 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
      241 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
      301 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
      361 ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact
      421 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
      481 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta
      541 ggcgtgtacg gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatccgct
      601 agcgctaccg gtcgccacca tggcctcctc cgaggacgtc atcaaggagt tcatgcgctt
      661 caaggtgcgc atggagggct ccgtgaacgg ccacgagttc gagatcgagg gcgagggcga
      721 gggccgcccc tacgagggca cccagaccgc caagctgaag gtgaccaagg gcggccccct
      781 gcccttcgcc tgggacatcc tgtcccctca gttccagtac ggctccaagg cctacgtgaa
      841 gcaccccgcc gacatccccg actacttgaa gctgtccttc cccgagggct tcaagtggga
      901 gcgcgtgatg aacttcgagg acggcggcgt ggtgaccgtg acccaggact cctccctgca
      961 ggacggcgag ttcatctaca aggtgaagct gcgcggcacc aacttcccct ccgacggccc
     1021 cgtaatgcag aagaagacca tgggctggga ggcctccacc gagcggatgt accccgagga
     1081 cggcgccctg aagggcgaga tcaagatgag gctgaagctg aaggacggcg gccactacga
     1141 cgccgaggtc aagaccacct acatggccaa gaagcccgtg cagctgcccg gcgcctacaa
     1201 gaccgacatc aagctggaca tcacctccca caacgaggac tacaccatcg tggaacagta
     1261 cgagcgcgcc gagggccgcc actccaccgg cgcctccgga ctcagatctc gagctcaagc
     1321 ttcgaattct atggcggtgt cggagagcca gctcaagaaa atggtgtcca agtacaaata
     1381 cagagaccta actgtacgtg aaactgtcaa tgttattact ctatacaaag atctcaaacc
     1441 tgttttggat tcatatgttt ttaacgatgg cagttccagg gaactaatga acctcactgg
     1501 aacaatccct gtgccttata gaggtaatac atacaatatt ccaatatgcc tatggctact
     1561 ggacacatac ccatataatc cccctatctg ttttgttaag cctactagtt caatgactat
     1621 taaaacagga aagcatgttg atgcaaatgg gaagatatat cttccttatc tacatgaatg
     1681 gaaacaccca cagtcagact tgttggggct tattcaggtc atgattgtgg tatttggaga
     1741 tgagccacct gtattttcac gaccgatatc ggcatcctat ccgccatacc aggcaacggg
     1801 gccaccaaat acttcctaca tgccaggcat gccaggtgga atctctccat acccatccgg
     1861 ataccctccc aatcccagtg gttacccagg ctgtccttac ccacctggtg gtccatatcc
     1921 tgccacaaca agttctcagt acccttctca gcctcctgtg accactgttg gtcccagtag
     1981 ggatggcaca atcagcgagg acaccatccg agcctctctc atctctgcgg tcagtgacaa
     2041 actgagatgg cggatgaagg aggaaatgga tcgtgcccag gcagagctca atgccttgaa
     2101 acgaacagaa gaagacctga aaaagggtca ccagaaactg gaagagatgg ttacccgttt
     2161 agatcaagaa gtagccgagg ttgataaaaa catagaactt ttgaaaaaga aggatgaaga
     2221 actcagttct gctctggaaa aaatggaaaa tcagtctgaa aacaatgata tcgatgaagt
     2281 tatcattccc acagctccct tatacaaaca gatcctgaat ctgtatgcag aagaaaacgc
     2341 tattgaagac actatctttt acttgggaga agccttgaga aggggcgtga tagacctgga
     2401 tgtcttcctg aagcatgtac gtcttctgtc ccgtaaacag ttccagctga gggcactaat
     2461 gcaaaaagca agaaagactg ccggtctcag tgacctctac tgaggatcca ccggatctag
     2521 ataactgatc ataatcagcc ataccacatt tgtagaggtt ttacttgctt taaaaaacct
     2581 cccacacctc cccctgaacc tgaaacataa aatgaatgca attgttgttg ttaacttgtt
     2641 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     2701 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttaacgcgt
     2761 aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat
     2821 tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga
     2881 tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca
     2941 acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct
     3001 aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc
     3061 cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag
     3121 cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca
     3181 cacccgccgc gcttaatgcg ccgctacagg gcgcgtcagg tggcactttt cggggaaatg
     3241 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     3301 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtcct gaggcggaaa
     3361 gaaccagctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg
     3421 cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg
     3481 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc
     3541 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
     3601 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt
     3661 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaagatc gatcaagaga
     3721 caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg
     3781 cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg
     3841 ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt
     3901 ccggtgccct gaatgaactg caagacgagg cagcgcggct atcgtggctg gccacgacgg
     3961 gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat
     4021 tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat
     4081 ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg
     4141 accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg
     4201 atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc
     4261 tcaaggcgag catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc
     4321 cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg
     4381 tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg
     4441 gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca
     4501 tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac
     4561 cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga
     4621 aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga
     4681 tctcatgctg gagttcttcg cccaccctag ggggaggcta actgaaacac ggaaggagac
     4741 aataccggaa ggaacccgcg ctatgacggc aataaaaaga cagaataaaa cgcacggtgt
     4801 tgggtcgttt gttcataaac gcggggttcg gtcccagggc tggcactctg tcgatacccc
     4861 accgagaccc cattggggcc aatacgcccg cgtttcttcc ttttccccac cccacccccc
     4921 aagttcgggt gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc
     4981 ctcaggttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
     5041 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     5101 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     5161 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     5221 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     5281 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     5341 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     5401 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     5461 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     5521 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     5581 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     5641 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     5701 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     5761 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
     5821 ggataaccgt attaccgc
//
