LOCUS       sequence-211653-        7417 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 7417)
  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 7417)
  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: Eps15-pmCherryN1
FEATURES             Location/Qualifiers
     source          1..7417
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        100..291
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     248..268
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(293..397)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      424..879
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(511..530)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     721..742
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    886..1007
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(897..916)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     951..970
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(1135..1839)
                     /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
                     EGRHSTGGMDELY"
     primer_bind     complement(1407..1426)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     1676..1694
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     regulatory      1836..1845
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(4644..4847)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(4673..4693)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        4848..5151
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(5326..5914)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5406..5425)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    6243..6290
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     6315..6334
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(6522..7316)
                     /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             complement(6522..7316)
                     /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(6633..6652)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     7243..7262
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(join(7351..7417,1..291))
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      join(7365..7417,1..83)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 gcctcggcct ctgcataaat aaaaaaaatt agtcagccat ggggcggaga atgggcggaa
       61 ctgggcggag ttaggggcgg gatgggcgga gttaggggcg ggactatggt tgctgactaa
      121 ttgagatgca tgctttgcat acttctgcct gctggggagc ctggggactt tccacacctg
      181 gttgctgact aattgagatg catgctttgc atacttctgc ctgctgggga gcctggggac
      241 tttccacacc ctaactgaca cacattccac agctggttct ttccgcctca ggactcttcc
      301 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
      361 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
      421 ctgacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
      481 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
      541 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
      601 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg
      661 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
      721 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt
      781 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
      841 ttaacgcgaa ttttaacaaa atattaacgc ttacaattta cgccttaaga tacattgatg
      901 agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg
      961 atgctattgc tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt
     1021 gcattcattt tatgtttcag gttcaggggg aggtgtggga ggttttttaa agcaagtaaa
     1081 acctctacaa atgtggtatg gctgattatg atctagagtc gcggccgctt tattgtacag
     1141 ctcgtccatg ccgccggtgg agtggcggcc ctcggcgcgt tcgtactgtt ccacgatggt
     1201 gtagtcctcg ttgtgggagg tgatgtccaa cttgatgttg acgttgtagg cgccgggcag
     1261 ctgcacgggc ttcttggcct tgtaggtggt cttgacctca gcgtcgtagt ggccgccgtc
     1321 cttcagcttc agcctctgct tgatctcgcc cttcagggcg ccgtcctcgg ggtacatccg
     1381 ctcggaggag gcctcccagc ccatggtctt cttctgcatt acggggccgt cggaggggaa
     1441 gttggtgccg cgcagcttca ccttgtagat gaactcgccg tcctgcaggg aggagtcctg
     1501 ggtcacggtc accacgccgc cgtcctcgaa gttcatcacg cgctcccact tgaagccctc
     1561 ggggaaggac agcttcaagt agtcggggat gtcggcgggg tgcttcacgt aggccttgga
     1621 gccgtacatg aactgagggg acaggatgtc ccaggcgaag ggcagggggc cacccttggt
     1681 caccttcagc ttggcggtct gggtgccctc gtaggggcgg ccctcgccct cgccctcgat
     1741 ctcgaactcg tggccgttca cggagccctc catgtgcacc ttgaagcgca tgaactcctt
     1801 gatgatggcc atgttatcct cctcgccctt gctcaccatg gtggcgaccg gtggatcccg
     1861 ggccagctga cccttggtcg aacccgaggt cgaacccatc cgtgcttctg atatctcaga
     1921 tttgctgagt gcaatagcca gttctaagtc ttcttgttcc tgctgattta gtcgggcaag
     1981 cctctgctct tcctctctct cactttccct cttggcccat tcgatcatat cttcttcaga
     2041 gggataagca ctgaagttgg caaaattgct tggatcagcc tctttattgg tagtggtagt
     2101 agcagaagtg aatggatcac aaaatatttc aggatctttt tctttggggc tatcgttgcc
     2161 tgggaaaggc tgaaatggat cattcagttt aaagggatca ggagaatcca atttgttgat
     2221 ggatcttttc ccaggtggta gagggcaggg tcttgttgga gttccgatct ttggtggcag
     2281 tgctgggggt tcatcttcac ttttgaccga tgtttcctca aatacatttt ttgtaatcac
     2341 tacgttgctg acagagctcg atgtggctga acgaaaagga tcttcattgt tgacctttga
     2401 caatgtgctg aagtcagcaa atccacctcc aaatgattca ttcccaaaaa cagaagcaaa
     2461 ggggtctgtg gctgaatcgc ttgctgcaac aactgttcca ccaggagcaa aaggatcatt
     2521 gtgcttcaac gtttctaccg atgtaatact gctattgttg gctgcactga aagggtcagt
     2581 gcttgaagtg gcaaaaggat cagtagattg cctgaagaaa cagtctgatg caaatggatc
     2641 tgaacctttg aaaggatcac caccaaatgg atcgattttt ccaaaaggat catccttgaa
     2701 aggatcactg ccaacaaaag gatcagactg gaaaaaatcc aagtttgtat ctgcaactgg
     2761 acctgtcagc gaacttgagt ctacattaaa tggatcttcc tcttttgaat gtctattatt
     2821 gtcgagttca gaacaaactt tttcagtaac agctgtagtc acctcatttt catcagtcac
     2881 accagaaggc agtagttcag gactacttct tgctggagat tcctggtgta tgggctcaga
     2941 ctctaggttg ctctggcctt caacatgttc attaaggttg gctgtttcac tgctgctggt
     3001 gctgaggctg caataatctg tagctccgtt tacaagaatg ctgtgtgggc tactgcacca
     3061 atttaactga ctattatgat tttccaaatc tttcatttcc atcagtttca tttgcattga
     3121 actaatttcc tgttgtgaat cttgtaggtg ctgctgaaga ggttccaact gagccttccc
     3181 tgactctaca ctctcctcca attctgctgt ttcttgctgt agacggctca gctcttctct
     3241 agcttttgcc aattcttctt cgtaagtgga gatctgcgat tcctgactag ttaattcagc
     3301 tttcagagaa gagatcagtt gggcctcctc agcacatttc tttctgactt ccttgagttg
     3361 ctcctccagc tgggctttct gctcatccag ttcatcaagg agttcctgta cctgctgttt
     3421 ctgggcctgt agtttttgca gattagtatt ctccctttga acttcatctt gaagatcctg
     3481 aacctcactt gtcctctgtt taatagtatc ttccttctcc ttaaggtcct gttccacatt
     3541 attcttttcc ctctgtaggt caactatttc attgttaaga gtatctagtt ccttaatagc
     3601 agagaaatct gcaacaggac ttgatcctat gatgttcttt tgtaaactgg ccctgtctga
     3661 tggtggaatc atttcaggag taagaacgtg aggaggatca atgcccttga ttaacttctg
     3721 actgattaag tgaaaagcca aggcaaactg atcctttgaa agcttcccac agtcctttgt
     3781 gtcgcataat gaccatatat gggctagtaa ggtagaaggt aaacctgttt tcaagaatat
     3841 ttcacggacc tccaatccag acacaaatcc gtccatatct ttatcagttt tcaggaagat
     3901 ttcatcatat ttagcttttt ctgcagggga tacaacccac gtttttctct tagatggtgg
     3961 caccaaggct ggaggcaagg acattggcac aggttctttc tccagtgcac agtataccaa
     4021 aaacatggca actgcaaact catctctgtc aagcattcca tcatggtcaa tatcactcaa
     4081 ctcccaaact cttccaagga tatccacagg taacttagag ttgagcaaca ctggtttcac
     4141 tttatcacca gacagaaatc cattcactgg gcttaaacta tcaaatattg catcatattt
     4201 ggccttatct tcaggtttta cagcccatgg gagctcagct gcagaggttc cactgattag
     4261 caaaggacta ctggtatcat gaaatcttgg tggaggaaca gccaggttca aactacttag
     4321 tgaaacttcc aatccattct gggcacatgc cacaagacgc aaagcaacaa agaattcttg
     4381 tttgttcagg atacctttgc catctgtgtc ggctaaatcc caaatctttc caagtatcaa
     4441 gtctggaagc cctgattttt tcaggaaagc agcagcatca gaagccaaca cccttccagt
     4501 attgcctgta tcaacctgtc tatagtattt ttcatataca ggattcccac ttgataactg
     4561 tgtcagggag agctgggccg ccgcagccat ggtctcgaga tctgagtccg gtagcgctag
     4621 cggatctgac ggttcactaa accagctctg cttatataga cctcccaccg tacacgccta
     4681 ccgcccattt gcgtcaatgg ggcggagttg ttacgacatt ttggaaagtc ccgttgattt
     4741 tggtgccaaa acaaactccc attgacgtca atggggtgga gacttggaaa tccccgtgag
     4801 tcaaaccgct atccacgccc attgatgtac tgccaaaacc gcatcaccat ggtaatagcg
     4861 atgactaata cgtagatgta ctgccaagta ggaaagtccc ataaggtcat gtactgggca
     4921 taatgccagg cgggccattt accgtcattg acgtcaatag ggggcgtact tggcatatga
     4981 tacacttgat gtactgccaa gtgggcagtt taccgtaaat actccaccca ttgacgtcaa
     5041 tggaaagtcc ctattggcgt tactatggga acatacgtca ttattgacgt caatgggcgg
     5101 gggtcgttgg gcggtcagcc aggcgggcca tttaccgtaa gttatgtaac gcggaactcc
     5161 atatatgggc tatgaactaa tgaccccgta attgattact attaataact aatgcatggc
     5221 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     5281 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     5341 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     5401 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     5461 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     5521 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     5581 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     5641 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     5701 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     5761 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     5821 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     5881 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     5941 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     6001 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     6061 atatgagtaa cctgaggcta tggcagggcc tgccgccccg acgttggctg cgagccctgg
     6121 gccttcaccc gaacttgggg ggtggggtgg ggaaaaggaa gaaacgcggg cgtattggcc
     6181 ccaatggggt ctcggtgggg tatcgacaga gtgccagccc tgggaccgaa ccccgcgttt
     6241 atgaacaaac gacccaacac cgtgcgtttt attctgtctt tttattgccg tcatagcgcg
     6301 ggttccttcc ggtattgtct ccttccgtgt ttcagttagc ctccccctag ggtgggcgaa
     6361 gaactccagc atgagatccc cgcgctggag gatcatccag ccggcgtccc ggaaaacgat
     6421 tccgaagccc aacctttcat agaaggcggc ggtggaatcg aaatctcgtg atggcaggtt
     6481 gggcgtcgct tggtcggtca tttcgaaccc cagagtcccg ctcagaagaa ctcgtcaaga
     6541 aggcgataga aggcgatgcg ctgcgaatcg ggagcggcga taccgtaaag cacgaggaag
     6601 cggtcagccc attcgccgcc aagctcttca gcaatatcac gggtagccaa cgctatgtcc
     6661 tgatagcggt ccgccacacc cagccggcca cagtcgatga atccagaaaa gcggccattt
     6721 tccaccatga tattcggcaa gcaggcatcg ccatgggtca cgacgagatc ctcgccgtcg
     6781 ggcatgctcg ccttgagcct ggcgaacagt tcggctggcg cgagcccctg atgctcttcg
     6841 tccagatcat cctgatcgac aagaccggct tccatccgag tacgtgctcg ctcgatgcga
     6901 tgtttcgctt ggtggtcgaa tgggcaggta gccggatcaa gcgtatgcag ccgccgcatt
     6961 gcatcagcca tgatggatac tttctcggca ggagcaaggt gagatgacag gagatcctgc
     7021 cccggcactt cgcccaatag cagccagtcc cttcccgctt cagtgacaac gtcgagcaca
     7081 gctgcgcaag gaacgcccgt cgtggccagc cacgatagcc gcgctgcctc gtcttgcagt
     7141 tcattcaggg caccggacag gtcggtcttg acaaaaagaa ccgggcgccc ctgcgctgac
     7201 agccggaaca cggcggcatc agagcagccg attgtctgtt gtgcccagtc atagccgaat
     7261 agcctctcca cccaagcggc cggagaacct gcgtgcaatc catcttgttc aatcatgcga
     7321 aacgatcctc atcctgtctc ttgatcgatc tttgcaaaag cctaggcctc caaaaaagcc
     7381 tcctcactac ttctggaata gctcagaggc cgaggcg
//
