LOCUS       sequence-148221-        5373 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 5373)
  AUTHORS   Choudhury A, Dominguez M, Puri V, Sharma DK, Narita K, Wheatley CL, 
            Marks DL, Pagano RE
  TITLE     Rab proteins mediate Golgi transport of caveola-internalized 
            glycosphingolipids and correct lipid trafficking in Niemann-Pick C 
            cells.
  JOURNAL   J Clin Invest. 2002 Jun . 109(12):1541-50.
  PUBMED    12070301
REFERENCE   2  (bases 1 to 5373)
  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: "J Clin 
            Invest. 2002 Jun . 109(12):1541-50."
COMMENT     Sequence Label: DsRed-rab7 DN
FEATURES             Location/Qualifiers
     source          1..5373
                     /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             join(620..622,623..625,626..1297)
                     /codon_start=1
                     /product="wild-type DsRed"
                     /label=DsRed1
                     /note="mammalian codon-optimized"
                     /translation="MVRSSKNVIKEFMRFKVRMEGTVNGHEFEIEGEGEGRPYEGHNTV
                     KLKVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     VVTVTQDSSLQDGCFIYKVKFIGVNFPSDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     KALKLKDGGHYLVEFKSIYMAKKPVQLPGYYYVDSKLDITSHNEDYTIVEQYERTEGRH
                     HLFL"
     primer_bind     complement(803..823)
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     primer_bind     1021..1040
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     1215..1238
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     misc_feature    1301..1357
                     /label=MCS
                     /note="multiple cloning site"
     CDS             1361..1387
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1388..1414
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    2168..2289
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2205..2224)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2259..2278
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2296..2751)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2433..2454)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2645..2664
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2778..2882
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2884..3241
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2884..3075
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2907..2927)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3092..3227
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3276..4070
                     /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             3276..4070
                     /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(3330..3349)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3940..3959
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4258..4277)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4302..4349
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4678..5266
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5167..5186
                     /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 tggtgcgctc ctccaagaac gtcatcaagg agttcatgcg
      661 cttcaaggtg cgcatggagg gcaccgtgaa cggccacgag ttcgagatcg agggcgaggg
      721 cgagggccgc ccctacgagg gccacaacac cgtgaagctg aaggtgacca agggcggccc
      781 cctgcccttc gcctgggaca tcctgtcccc ccagttccag tacggctcca aggtgtacgt
      841 gaagcacccc gccgacatcc ccgactacaa gaagctgtcc ttccccgagg gcttcaagtg
      901 ggagcgcgtg atgaacttcg aggacggcgg cgtggtgacc gtgacccagg actcctccct
      961 gcaggacggc tgcttcatct acaaggtgaa gttcatcggc gtgaacttcc cctccgacgg
     1021 ccccgtaatg cagaagaaga ccatgggctg ggaggcctcc accgagcgcc tgtacccccg
     1081 cgacggcgtg ctgaagggcg agatccacaa ggccctgaag ctgaaggacg gcggccacta
     1141 cctggtggag ttcaagtcca tctacatggc caagaagccc gtgcagctgc ccggctacta
     1201 ctacgtggac tccaagctgg acatcacctc ccacaacgag gactacacca tcgtggagca
     1261 gtacgagcgc accgagggcc gccaccacct gttcctgctc agatctcgag ctcaagcttc
     1321 gaattctgca gtcgacggta ccgcgggccc gggatccatg tacccatacg atgttccgga
     1381 ttacgcttac ccatacgatg ttccggatta cgctggtacc tctaggaaga aagtgttgct
     1441 gaaggttatc atcctgggag attctggagt cgggaagaac tcactcatga accagtatgt
     1501 gaataagaaa ttcagcaatc agtacaaagc cacaatagga gctgactttc tgaccaagga
     1561 ggtgatggtg gatgacaggc tagtcacaat gcagatatgg gacacagcag gacaggaacg
     1621 gttccagtct ctcggtgtgg ccttctacag aggtgcagac tgctgcgttc tggtatttga
     1681 tgtgactgcc cccaacacat tcaaaaccct agatagctgg agagatgagt ttctcatcca
     1741 ggccagtccc cgagatcctg aaaacttccc atttgttgtg ttgggaaaca agattgacct
     1801 cgaaaacaga caagtggcca caaagcgggc acaggcctgg tgctacagca aaaacaacat
     1861 tccctacttt gagaccagtg ccaaggaggc catcaacgtg gagcaggcgt tccagacgat
     1921 tgcacggaat gcacttaagc aggaaacgga ggtggagctg tacaacgaat ttcctgaacc
     1981 tatcaaactg gacaagaatg accgggccaa ggcctcggca gaaagctgca gttgctgagg
     2041 atccaccgga tctagataac tgatcataat cagccatacc acatttgtag aggttttact
     2101 tgctttaaaa aacctcccac acctccccct gaacctgaaa cataaaatga atgcaattgt
     2161 tgttgttaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     2221 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     2281 tgtatcttaa cgcgtaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt
     2341 gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa
     2401 aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa
     2461 agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac
     2521 gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga
     2581 accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa
     2641 aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc
     2701 tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg tcaggtggca
     2761 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     2821 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     2881 gtcctgaggc ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc
     2941 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg
     3001 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     3061 agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc
     3121 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
     3181 ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa
     3241 agatcgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg
     3301 caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa
     3361 tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg
     3421 tcaagaccga cctgtccggt gccctgaatg aactgcaaga cgaggcagcg cggctatcgt
     3481 ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa
     3541 gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc
     3601 ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg
     3661 ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg
     3721 aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg
     3781 aactgttcgc caggctcaag gcgagcatgc ccgacggcga ggatctcgtc gtgacccatg
     3841 gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact
     3901 gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg
     3961 ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc
     4021 ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct
     4081 ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac
     4141 cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat
     4201 cctccagcgc ggggatctca tgctggagtt cttcgcccac cctaggggga ggctaactga
     4261 aacacggaag gagacaatac cggaaggaac ccgcgctatg acggcaataa aaagacagaa
     4321 taaaacgcac ggtgttgggt cgtttgttca taaacgcggg gttcggtccc agggctggca
     4381 ctctgtcgat accccaccga gaccccattg gggccaatac gcccgcgttt cttccttttc
     4441 cccaccccac cccccaagtt cgggtgaagg cccagggctc gcagccaacg tcggggcggc
     4501 aggccctgcc atagcctcag gttactcata tatactttag attgatttaa aacttcattt
     4561 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     4621 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     4681 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     4741 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     4801 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     4861 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     4921 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     4981 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     5041 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     5101 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     5161 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     5221 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     5281 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     5341 atcccctgat tctgtggata accgtattac cgc
//
