LOCUS       sequence-176426-        5369 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 5369)
  AUTHORS   Sharma DK, Choudhury A, Singh RD, Wheatley CL, Marks DL, Pagano RE
  TITLE     Glycosphingolipids internalized via caveolar-related endocytosis 
            rapidly merge with the clathrin pathway in early endosomes and form 
            microdomains for recycling.
  JOURNAL   J Biol Chem. 2003 Feb 28. 278(9):7564-72.
  PUBMED    12482757
REFERENCE   2  (bases 1 to 5369)
  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 Biol 
            Chem. 2003 Feb 28. 278(9):7564-72."
COMMENT     Sequence Label: DsRed-Rab5 DN
FEATURES             Location/Qualifiers
     source          1..5369
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        97..400
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        401..604
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     555..575
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      643..652
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             649..1323
                     /codon_start=1
                     /product="improved tetrameric variant of DsRed fluorescent 
                     protein"
                     /label=DsRed2
                     /note="mammalian codon-optimized"
                     /translation="MASSENVITEFMRFKVRMEGTVNGHEFEIEGEGEGRPYEGHNTVK
                     LKVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGV
                     ATVTQDSSLQDGCFIYKVKFIGVNFPSDGPVMQKKTMGWEASTERLYPRDGVLKGETHK
                     ALKLKDGGHYLVEFKSIYMAKKPVQLPGYYYVDAKLDITSHNEDYTIVEQYERTEGRHH
                     LFL"
     primer_bind     complement(829..849)
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     primer_bind     1241..1264
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     regulatory      1330..1339
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1342..1368
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     misc_feature    2017..2069
                     /label=MCS
                     /note="multiple cloning site"
     polyA_signal    2193..2314
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2230..2249)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2284..2303
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2321..2776)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2458..2479)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2670..2689
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2803..2907
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2909..3266
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2909..3100
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2932..2952)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3117..3252
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3301..4095
                     /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             3301..4095
                     /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(3355..3374)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3965..3984
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4283..4302)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4327..4374
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4703..5291
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5192..5211
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgattctg tggataaccg tattaccgcc atgcattagt tattaatagt aatcaattac
       61 ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg
      121 cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc
      181 catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac
      241 tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa
      301 tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac
      361 ttggcagtac atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta
      421 catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga
      481 cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa
      541 ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag
      601 agctggttta gtgaaccgtc agatccgcta gcgctaccgg tcgccaccat ggcctcctcc
      661 gagaacgtca tcaccgagtt catgcgcttc aaggtgcgca tggagggcac cgtgaacggc
      721 cacgagttcg agatcgaggg cgagggcgag ggccgcccct acgagggcca caacaccgtg
      781 aagctgaagg tgaccaaggg cggccccctg cccttcgcct gggacatcct gtccccccag
      841 ttccagtacg gctccaaggt gtacgtgaag caccccgccg acatccccga ctacaagaag
      901 ctgtccttcc ccgagggctt caagtgggag cgcgtgatga acttcgagga cggcggcgtg
      961 gcgaccgtga cccaggactc ctccctgcag gacggctgct tcatctacaa ggtgaagttc
     1021 atcggcgtga acttcccctc cgacggcccc gtgatgcaga agaagaccat gggctgggag
     1081 gcctccaccg agcgcctgta cccccgcgac ggcgtgctga agggcgagac ccacaaggcc
     1141 ctgaagctga aggacggcgg ccactacctg gtggagttca agtccatcta catggccaag
     1201 aagcccgtgc agctgcccgg ctactactac gtggacgcca agctggacat cacctcccac
     1261 aacgaggact acaccatcgt ggagcagtac gagcgcaccg agggccgcca ccacctgttc
     1321 ctgagatctg ccgccatggc atatccatat gacgtcccag actatgctgg cgctagtcga
     1381 ggcgcaacaa gacccaacgg accaaatact ggaaataaaa tatgccagtt caaactagta
     1441 cttctgggag agtctgctgt tggcaaatca agcctagtgc ttcgttttgt gaagggccaa
     1501 tttcatgaat ttcaagagag taccataggg gctgcttttc taacccaaac tgtgtgtctt
     1561 gatgatacaa cagtaaagtt tgaaatatgg gatacagctg gtcaagaacg ataccatagc
     1621 ttagcaccaa tgtactacag aggagcacaa gcagccatag ttgtatatga tatcacaaat
     1681 gaggagtcct ttgccagagc caaaaactgg gttaaagaac ttcagaggca agccagtcct
     1741 aacattgtaa tagctttatc aggaatcaag gctgatcttg caaataaaag agctgtcgat
     1801 ttccaggaag cacagtccta tgcagatgac aacagtttat tattcatgga gacatcagct
     1861 aaaacatcga tgaatgtaaa tgaaatattc atggcaatag ctaaaaagtt gccaaagaac
     1921 gaaccacaga atccaggagc aaattctgcc agaggaagag gagtagacct tactgaaccc
     1981 acgcagccaa ccaggaatca gtgttgtagt aactagctcg agctcaagct tcgaattctg
     2041 cagtcgacgg taccgcgggc ccgggatcca ccggatctag ataactgatc ataatcagcc
     2101 ataccacatt tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc
     2161 tgaaacataa aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt
     2221 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     2281 gttgtggttt gtccaaactc atcaatgtat cttaacgcgt aaattgtaag cgttaatatt
     2341 ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa
     2401 atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca
     2461 gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc
     2521 gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg
     2581 aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg
     2641 ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg
     2701 gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg
     2761 ccgctacagg gcgcgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     2821 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     2881 cttcaataat attgaaaaag gaagagtcct gaggcggaaa gaaccagctg tggaatgtgt
     2941 gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     3001 atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta
     3061 tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc
     3121 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
     3181 tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct
     3241 tttttggagg cctaggcttt tgcaaagatc gatcaagaga caggatgagg atcgtttcgc
     3301 atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc
     3361 ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca
     3421 gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg
     3481 caagacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg
     3541 ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag
     3601 gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg
     3661 cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc
     3721 atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa
     3781 gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgag catgcccgac
     3841 ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat
     3901 ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac
     3961 atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc
     4021 ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt
     4081 gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc
     4141 tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg
     4201 ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg
     4261 cccaccctag ggggaggcta actgaaacac ggaaggagac aataccggaa ggaacccgcg
     4321 ctatgacggc aataaaaaga cagaataaaa cgcacggtgt tgggtcgttt gttcataaac
     4381 gcggggttcg gtcccagggc tggcactctg tcgatacccc accgagaccc cattggggcc
     4441 aatacgcccg cgtttcttcc ttttccccac cccacccccc aagttcgggt gaaggcccag
     4501 ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc ctcaggttac tcatatatac
     4561 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     4621 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     4681 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     4741 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     4801 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     4861 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     4921 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     4981 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     5041 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     5101 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     5161 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     5221 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     5281 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     5341 ttgctcacat gttctttcct gcgttatcc
//
