LOCUS       sequence-157882-        6030 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 6030)
  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 6030)
  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-rab9 WT
FEATURES             Location/Qualifiers
     source          1..6030
                     /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"
     CDS             2033..2059
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2060..2086
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    2825..2946
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2862..2881)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2916..2935
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2953..3408)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3090..3111)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3302..3321
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3435..3539
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3541..3898
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3541..3732
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3564..3584)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3749..3884
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3933..4727
                     /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             3933..4727
                     /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(3987..4006)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4597..4616
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4915..4934)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4959..5006
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5335..5923
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5824..5843
                     /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 gcaggaaaat catcactttt
     1441 taaagtaatt ctccttggag atggtggagt tgggaagagt tcacttatga acagatatgt
     1501 aactaataag tttgataccc agctcttcca tacaataggt gtggaatttt taaataaaga
     1561 tttggaagtg gatggacatt ttgttaccat gcagatttgg gacacggcag gtcaggagcg
     1621 attccgaagc ctgaggacac cattttacag aggttctgac tgctgcctgc ttacttttag
     1681 tgtcgatgat tcacaaagct tccagaactt aagtaactgg aagaaagaat tcatatatta
     1741 tgcagatgtg aaagagcctg agagctttcc ttttgtgatt ctgggtaaca agattgacat
     1801 aagcgaacgg caggtgtcta cagaagaagc ccaagcttgg tgcagggaca acggcgacta
     1861 tccttatttt gaaacaagtg caaaagatgc cacaaatgtg gcagcagcct ttgaggaagc
     1921 ggttcgaaga gttcttgcta ccgaggatag gtcagatcat ttgattcaga cagacacagt
     1981 caatcttcac cgaaagccca agcctagctc atcttgctgt tgaggatcca tgtacccata
     2041 cgatgttccg gattacgctt acccatacga tgttccggat tacgctggta ccgcaggaaa
     2101 atcatcactt tttaaagtaa ttctccttgg agatggtgga gttgggaaga gttcacttat
     2161 gaacagatat gtaactaata agtttgatac ccagctcttc catacaatag gtgtggaatt
     2221 tttaaataaa gatttggaag tggatggaca ttttgttacc atgcagattt gggacacggc
     2281 aggtcaggag cgattccgaa gcctgaggac accattttac agaggttctg actgctgcct
     2341 gcttactttt agtgtcgatg attcacaaag cttccagaac ttaagtaact ggaagaaaga
     2401 attcatatat tatgcagatg tgaaagagcc tgagagcttt ccttttgtga ttctgggtaa
     2461 caagattgac ataagcgaac ggcaggtgtc tacagaagaa gcccaagctt ggtgcaggga
     2521 caacggcgac tatccttatt ttgaaacaag tgcaaaagat gccacaaatg tggcagcagc
     2581 ctttgaggaa gcggttcgaa gagttcttgc taccgaggat aggtcagatc atttgattca
     2641 gacagacaca gtcaatcttc accgaaagcc caagcctagc tcatcttgct gttgaggatc
     2701 caccggatct agataactga tcataatcag ccataccaca tttgtagagg ttttacttgc
     2761 tttaaaaaac ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt
     2821 tgttaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     2881 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     2941 atcttaacgc gtaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt
     3001 aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag
     3061 aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga
     3121 acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg
     3181 aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc
     3241 ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg
     3301 aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc
     3361 gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt
     3421 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     3481 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagtc
     3541 ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg
     3601 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg
     3661 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     3721 aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca
     3781 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc
     3841 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaga
     3901 tcgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag
     3961 gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg
     4021 gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca
     4081 agaccgacct gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc
     4141 tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg
     4201 actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg
     4261 ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta
     4321 cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag
     4381 ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac
     4441 tgttcgccag gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg acccatggcg
     4501 atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg
     4561 gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg
     4621 aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg
     4681 attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg
     4741 gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc
     4801 cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct
     4861 ccagcgcggg gatctcatgc tggagttctt cgcccaccct agggggaggc taactgaaac
     4921 acggaaggag acaataccgg aaggaacccg cgctatgacg gcaataaaaa gacagaataa
     4981 aacgcacggt gttgggtcgt ttgttcataa acgcggggtt cggtcccagg gctggcactc
     5041 tgtcgatacc ccaccgagac cccattgggg ccaatacgcc cgcgtttctt ccttttcccc
     5101 accccacccc ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg gggcggcagg
     5161 ccctgccata gcctcaggtt actcatatat actttagatt gatttaaaac ttcattttta
     5221 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     5281 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     5341 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     5401 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     5461 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     5521 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     5581 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     5641 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     5701 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     5761 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     5821 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     5881 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     5941 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
     6001 ccctgattct gtggataacc gtattaccgc
//
