LOCUS       sequence-199003-        5358 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 5358)
  AUTHORS   Taylor MJ, Perrais D, Merrifield CJ
  TITLE     A high precision survey of the molecular dynamics of Mammalian 
            clathrin-mediated endocytosis.
  JOURNAL   PLoS Biol. 2011 Mar . 9(3):e1000604.
  PUBMED    21445324
REFERENCE   2  (bases 1 to 5358)
  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: "PLoS Biol. 
            2011 Mar . 9(3):e1000604."
COMMENT     Sequence Label: Rab5a-pmCherryC1
FEATURES             Location/Qualifiers
     source          1..5358
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(272..291)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    316..363
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      692..1280
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1181..1200
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        1455..1758
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1759..1962
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1913..1933
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             1994..2701
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(2139..2157)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2407..2426
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    3540..3661
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3577..3596)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3631..3650
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3668..4123)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3805..3826)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4017..4036
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4150..4254
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4256..4613
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4256..4447
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4279..4299)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4464..4599
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             join(4648..5358,1..84)
                     /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             join(4648..5358,1..84)
                     /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(4702..4721)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5312..5331
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
ORIGIN
        1 gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat
       61 cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga
      121 cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct
      181 tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg
      241 agttcttcgc ccaccctagg gggaggctaa ctgaaacacg gaaggagaca ataccggaag
      301 gaacccgcgc tatgacggca ataaaaagac agaataaaac gcacggtgtt gggtcgtttg
      361 ttcataaacg cggggttcgg tcccagggct ggcactctgt cgatacccca ccgagacccc
      421 attggggcca atacgcccgc gtttcttcct tttccccacc ccacccccca agttcgggtg
      481 aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc tgccatagcc tcaggttact
      541 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
      601 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
      661 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
      721 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
      781 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
      841 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
      901 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
      961 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     1021 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     1081 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     1141 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     1201 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     1261 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     1321 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     1381 ttaccgccat gcattagtta ttaatagtaa tcaattacgg ggtcattagt tcatagccca
     1441 tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac
     1501 gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact
     1561 ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa
     1621 gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg
     1681 cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta
     1741 gtcatcgcta ttaccatggt gatgcggttt tggcagtaca tcaatgggcg tggatagcgg
     1801 tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg
     1861 caccaaaatc aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg
     1921 ggcggtaggc gtgtacggtg ggaggtctat ataagcagag ctggtttagt gaaccgtcag
     1981 atccgctagc accatggtga gcaagggcga ggaggataac atggccatca tcaaggagtt
     2041 catgcgcttc aaggtgcaca tggagggctc cgtgaacggc cacgagttcg agatcgaggg
     2101 cgagggcgag ggccgcccct acgagggcac ccagaccgcc aagctgaagg tgaccaaggg
     2161 tggccccctg cccttcgcct gggacatcct gtcccctcag ttcatgtacg gctccaaggc
     2221 ctacgtgaag caccccgccg acatccccga ctacttgaag ctgtccttcc ccgagggctt
     2281 caagtgggag cgcgtgatga acttcgagga cggcggcgtg gtgaccgtga cccaggactc
     2341 ctccctgcag gacggcgagt tcatctacaa ggtgaagctg cgcggcacca acttcccctc
     2401 cgacggcccc gtaatgcaga agaagaccat gggctgggag gcctcctccg agcggatgta
     2461 ccccgaggac ggcgccctga agggcgagat caagcagagg ctgaagctga aggacggcgg
     2521 ccactacgac gctgaggtca agaccaccta caaggccaag aagcccgtgc agctgcccgg
     2581 cgcctacaac gtcaacatca agttggacat cacctcccac aacgaggact acaccatcgt
     2641 ggaacagtac gaacgcgccg agggccgcca ctccaccggc ggcatggacg agctgtacaa
     2701 gtccggactc agatctggtt caggatctgg aatggctaat cgaggagcaa caagacccaa
     2761 cgggccaaat actggaaata aaatatgcca gttcaaactg gtccttctag gagagtctgc
     2821 tgttggcaaa tcaagcctgg ttcttcgctt tgtgaaaggc caatttcatg aatttcaaga
     2881 gagtaccatt ggggctgcct ttctaaccca aactgtgtgt cttgatgaca caacagtaaa
     2941 atttgaaata tgggatacag ctggtcaaga acggtatcat agcttagcac caatgtacta
     3001 ccgaggagca caagcagcca tagttgtgta tgatatcaca aatgaggaat cctttgcgag
     3061 agcaaaaaac tgggttaaag aacttcaaag gcaagcaagt cctaatattg tgatagcttt
     3121 gtcaggaaac aaagctgact tagcaaataa aagagctgtt gacttccagg aagcacagtc
     3181 ctatgcagat gacaacagct tattatttat ggagacatca gctaagacat caatgaatgt
     3241 aaatgaaata tttatggcaa tagctaaaaa gctgccaaag aatgaaccac agaatcctgg
     3301 tgcaaactca gccagaggac gaggagtaga ccttactgag cctgcacagc cagccagaag
     3361 ccagtgttgt agtaactgag aattctgcag tcgacggtac cgcgggcccg ggatccaccg
     3421 gatctagata actgatcata atcagccata ccacatttgt agaggtttta cttgctttaa
     3481 aaaacctccc acacctcccc ctgaacctga aacataaaat gaatgcaatt gttgttgtta
     3541 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     3601 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     3661 aacgcgtaaa ttgtaagcgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc
     3721 agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag
     3781 accgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg
     3841 gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca
     3901 tcaccctaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa
     3961 gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg
     4021 aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta
     4081 accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtcaggtgg cacttttcgg
     4141 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     4201 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtcctgag
     4261 gcggaaagaa ccagctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc
     4321 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt
     4381 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     4441 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
     4501 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg
     4561 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaagatcgat
     4621 caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct
     4681 ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc
     4741 tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc
     4801 gacctgtccg gtgccctgaa tgaactgcaa gacgaggcag cgcggctatc gtggctggcc
     4861 acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg
     4921 ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag
     4981 aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc
     5041 ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt
     5101 cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc
     5161 gccaggctca aggcgagcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc
     5221 tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg
     5281 ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag
     5341 cttggcggcg aatgggct
//
