LOCUS       sequence-169555-        7273 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 7273)
  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 7273)
  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-hALIX
FEATURES             Location/Qualifiers
     source          1..7273
                     /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    4068..4189
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4105..4124)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4159..4178
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4196..4651)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4333..4354)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4545..4564
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4678..4782
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4784..5141
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4784..4975
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4807..4827)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4992..5127
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5176..5970
                     /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             5176..5970
                     /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(5230..5249)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5840..5859
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6158..6177)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6202..6249
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6578..7166
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7067..7086
                     /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 atggcgacat tcatctcggt gcagctgaaa aagacctcag aggtggacct
     1381 ggccaagccg ctggtgaagt tcatccagca gacttaccca agcggcgggg aagagcaggc
     1441 ccagtactgc cgcgcggcgg aggagctcag caagctgcgc cgcgccgcag tcggtcgtcc
     1501 gctggacaag cacgagggcg cgctcgagac gctcctgaga tattatgatc agatttgttc
     1561 tattgaaccc aaattcccat tttctgaaaa tcagatctgc ttgacattta cctggaagga
     1621 tgctttcgat aaaggttcac tttttggagg ctctgtaaaa ctggctcttg caagcttagg
     1681 atatgaaaag agctgtgtgt tgttcaattg tgcagcctta gctagccaaa ttgcagcaga
     1741 acagaacctg gataatgatg aaggattgaa aatcgctgct aaacattacc agtttgctag
     1801 tggtgccttt ttacatatta aagagacggt tttatctgcc ttaagtcgag agccgaccgt
     1861 ggacatatct ccagatactg ttgggaccct cagtcttatt atgctggcac aggctcaaga
     1921 agtatttttt ttaaaagcca caagagataa aatgaaagat gccatcatag ctaaattggc
     1981 taatcaggct gcagattatt ttggtgatgc tttcaaacag tgtcaataca aagatactct
     2041 ccccaaggag gtgttccctg tcttggctgc aaagcactgt atcatgcagg ccaatgctga
     2101 gtaccatcag tctatcctgg caaaacagca gaagaaattt ggagaagaaa ttgcaaggtt
     2161 acagcatgca gcagaactga ttaaaacagt ggcatctcgc tatgatgaat atgttaatgt
     2221 gaaggatttt tctgacaaaa tcaatcgtgc ccttgctgca gcaaagaagg ataatgactt
     2281 catttatcat gatcgagttc cagaccttaa agatctagat cctattggca aagccacact
     2341 tgtgaaatct accccggtca atgtacccat cagtcagaaa tttactgatc tgtttgagaa
     2401 gatggttccc gtgtcagtac agcagtcttt ggctgcctat aatcagagga aagccgattt
     2461 ggttaacaga tcaattgctc agatgagaga agccaccact ttggcaaatg gggtgctagc
     2521 ttcccttaat cttccagcag caattgaaga tgtgtctgga gacactgtac ctcagtctat
     2581 attgactaaa tccagatctg tgattgaaca gggaggcatc cagactgttg atcagttgat
     2641 taaagaactg cctgaattac tgcaacgaaa tagagaaatc ctagatgagt cattaaggtt
     2701 gttggatgaa gaagaagcaa ccgataatga tttaagagca aaatttaagg aacgttggca
     2761 aaggacacca tccaatgaac tgtataagcc tttaagagca gagggaacca acttcagaac
     2821 agttttagat aaagctgtgc aggcagatgg acaagtgaaa gaatgttacc agtctcatcg
     2881 tgacaccatc gtgcttttgt gtaagccaga gcctgagctg aatgctgcca tcccttctgc
     2941 taatccggca aagaccatgc agggcagtga ggttgtaaat gtcttaaaat ccttattgtc
     3001 aaatcttgat gaagtaaaga aggaaagaga gggtctggag aatgacttga aatctgtgaa
     3061 ttttgacatg acaagcaagt ttttgacagc cctggctcaa gatggtgtga taaatgaaga
     3121 agctctttct gttactgaac tagatcgagt ctatggaggt cttacaacta aagtccaaga
     3181 atctctaaag aaacaggagg gacttcttaa aaatattcag gtctcacatc aggaattttc
     3241 aaaaatgaaa caatctaata atgaagctaa cttaagagaa gaagttttga agaatttagc
     3301 tactgcatat gacaactttg ttgaacttgt agctaatttg aaggaaggca caaagtttta
     3361 caatgagttg actgaaatcc tggtcaggtt ccagaacaaa tgcagtgata tagtttttgc
     3421 acggaagaca gaaagagatg aactcttaaa ggacttgcaa caaagcattg ccagagaacc
     3481 tagtgctcct tcaattccta cacctgcgta tcagtcctca ccagcaggag gacatgcacc
     3541 aactcctcca actccagcgc caagaaccat gccgcctact aagccccagc ccccagccag
     3601 gcctccacca cctgtgcttc cagcaaatcg agctccttct gctactgctc catctccagt
     3661 gggggctggg actgctgcgc cagctccatc acaaacgcct ggctcagctc ctcctccaca
     3721 ggcgcaggga ccaccctatc ccacctatcc aggatatcct gggtattgcc aaatgcccat
     3781 gcccatgggc tataatcctt atgcgtatgg ccagtataat atgccatatc caccagtgta
     3841 tcaccagagt cctggacagg ctccataccc gggaccccag cagccttcat accccttccc
     3901 tcagccccca cagcagtctt actatccaca gccagtaagg atccaccgga tctagataac
     3961 tgatcataat cagccatacc acatttgtag aggttttact tgctttaaaa aacctcccac
     4021 acctccccct gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg
     4081 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     4141 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttaa cgcgtaaatt
     4201 gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt
     4261 aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg
     4321 ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc
     4381 aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca
     4441 agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga
     4501 tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa
     4561 ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc
     4621 gccgcgctta atgcgccgct acagggcgcg tcaggtggca cttttcgggg aaatgtgcgc
     4681 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     4741 taaccctgat aaatgcttca ataatattga aaaaggaaga gtcctgaggc ggaaagaacc
     4801 agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa
     4861 gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc
     4921 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc
     4981 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct
     5041 gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga
     5101 agtagtgagg aggctttttt ggaggcctag gcttttgcaa agatcgatca agagacagga
     5161 tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg
     5221 gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc
     5281 gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt
     5341 gccctgaatg aactgcaaga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt
     5401 ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc
     5461 gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc
     5521 atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac
     5581 caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag
     5641 gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag
     5701 gcgagcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat
     5761 atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg
     5821 gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa
     5881 tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc
     5941 ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc
     6001 aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt
     6061 tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca
     6121 tgctggagtt cttcgcccac cctaggggga ggctaactga aacacggaag gagacaatac
     6181 cggaaggaac ccgcgctatg acggcaataa aaagacagaa taaaacgcac ggtgttgggt
     6241 cgtttgttca taaacgcggg gttcggtccc agggctggca ctctgtcgat accccaccga
     6301 gaccccattg gggccaatac gcccgcgttt cttccttttc cccaccccac cccccaagtt
     6361 cgggtgaagg cccagggctc gcagccaacg tcggggcggc aggccctgcc atagcctcag
     6421 gttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     6481 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     6541 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     6601 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     6661 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     6721 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     6781 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     6841 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     6901 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     6961 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     7021 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     7081 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     7141 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     7201 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
     7261 accgtattac cgc
//
