LOCUS       sequence-194027-        5357 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 5357)
  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 5357)
  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: CLC-pmCherryC1
FEATURES             Location/Qualifiers
     source          1..5357
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        20..211
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     168..188
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(213..317)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      344..799
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(431..450)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     641..662
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    806..927
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(817..836)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     871..890
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(1765..2472)
                     /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(2040..2059)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     2309..2327
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     promoter        complement(2504..2707)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2533..2553)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2708..3011
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(3186..3774)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3266..3285)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4103..4150
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     4175..4194
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(4382..5176)
                     /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(4382..5176)
                     /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(4493..4512)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5103..5122
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(join(5211..5357,1..211))
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      join(5225..5357,1..3)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 gatgggcgga gttaggggcg ggactatggt tgctgactaa ttgagatgca tgctttgcat
       61 acttctgcct gctggggagc ctggggactt tccacacctg gttgctgact aattgagatg
      121 catgctttgc atacttctgc ctgctgggga gcctggggac tttccacacc ctaactgaca
      181 cacattccac agctggttct ttccgcctca ggactcttcc tttttcaata ttattgaagc
      241 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
      301 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgcgcc ctgtagcggc
      361 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
      421 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
      481 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
      541 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
      601 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
      661 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt
      721 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa
      781 atattaacgc ttacaattta cgcgttaaga tacattgatg agtttggaca aaccacaact
      841 agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta
      901 accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt tatgtttcag
      961 gttcaggggg aggtgtggga ggttttttaa agcaagtaaa acctctacaa atgtggtatg
     1021 gctgattatg atcagttatc tagatccggt ggatcccggg cccgcggtac ctcagtgcac
     1081 caggggggcc tgcttcaggg agatgaggac tgagcgcatg cgagagacat ctttggcctg
     1141 tttgctggac ttggggttaa agtcacacag gcgggccacc cgctcccact cagtgcctgg
     1201 ggatgactcg tcaatgtcat ttacaaaggc ttcttctgct gccctgttgt ttgcctttgt
     1261 cttctgtagc tgctcgtcct gcctcgcata ccactcttcc agctccttta tcgccttttc
     1321 tttccactcc gcttcttgct tccgagaatt ggcatcgagg gcttccaggc gctctgtctg
     1381 ctcttctctc cacttacgga tactttcagg ctctgattgc aatcgatcca cttctgaaat
     1441 ggctgcgtaa ctgtctgttg gaccattgct ctcctggtag tattcaccat tcatcactcc
     1501 gtcaacagca tccggacctc ccggcggctc gccgtgtggc tgcggcccgg gggcgccgcc
     1561 gtccaggatg gcgaaggcct catcgttctc gatgcccgca atctcactct cttgctgcgc
     1621 caagaaggcc gcggccgggt cttcttcgcc ggcgcccgcc accccgtttc ctagagcggg
     1681 gccaccgggc gcaccggcgg gagcgccgaa tggatccaac tcggccatag aattcgaagc
     1741 ttgagctcga gatctgagtc cggacttgta cagctcgtcc atgccgccgg tggagtggcg
     1801 gccctcggcg cgttcgtact gttccacgat ggtgtagtcc tcgttgtggg aggtgatgtc
     1861 caacttgatg ttgacgttgt aggcgccggg cagctgcacg ggcttcttgg ccttgtaggt
     1921 ggtcttgacc tcagcgtcgt agtggccgcc gtccttcagc ttcagcctct gcttgatctc
     1981 gcccttcagg gcgccgtcct cggggtacat ccgctcggag gaggcctccc agcccatggt
     2041 cttcttctgc attacggggc cgtcggaggg gaagttggtg ccgcgcagct tcaccttgta
     2101 gatgaactcg ccgtcctgca gggaggagtc ctgggtcacg gtcaccacgc cgccgtcctc
     2161 gaagttcatc acgcgctccc acttgaagcc ctcggggaag gacagcttca agtagtcggg
     2221 gatgtcggcg gggtgcttca cgtaggcctt ggagccgtac atgaactgag gggacaggat
     2281 gtcccaggcg aagggcaggg ggccaccctt ggtcaccttc agcttggcgg tctgggtgcc
     2341 ctcgtagggg cggccctcgc cctcgccctc gatctcgaac tcgtggccgt tcacggagcc
     2401 ctccatgtgc accttgaagc gcatgaactc cttgatgatg gccatgttat cctcctcgcc
     2461 cttgctcacc atggtgctag cggatctgac ggttcactaa accagctctg cttatataga
     2521 cctcccaccg tacacgccta ccgcccattt gcgtcaatgg ggcggagttg ttacgacatt
     2581 ttggaaagtc ccgttgattt tggtgccaaa acaaactccc attgacgtca atggggtgga
     2641 gacttggaaa tccccgtgag tcaaaccgct atccacgccc attgatgtac tgccaaaacc
     2701 gcatcaccat ggtaatagcg atgactaata cgtagatgta ctgccaagta ggaaagtccc
     2761 ataaggtcat gtactgggca taatgccagg cgggccattt accgtcattg acgtcaatag
     2821 ggggcgtact tggcatatga tacacttgat gtactgccaa gtgggcagtt taccgtaaat
     2881 actccaccca ttgacgtcaa tggaaagtcc ctattggcgt tactatggga acatacgtca
     2941 ttattgacgt caatgggcgg gggtcgttgg gcggtcagcc aggcgggcca tttaccgtaa
     3001 gttatgtaac gcggaactcc atatatgggc tatgaactaa tgaccccgta attgattact
     3061 attaataact aatgcatggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     3121 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     3181 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     3241 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     3301 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     3361 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     3421 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     3481 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     3541 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     3601 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     3661 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     3721 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     3781 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     3841 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     3901 tttaaatcaa tctaaagtat atatgagtaa cctgaggcta tggcagggcc tgccgccccg
     3961 acgttggctg cgagccctgg gccttcaccc gaacttgggg ggtggggtgg ggaaaaggaa
     4021 gaaacgcggg cgtattggcc ccaatggggt ctcggtgggg tatcgacaga gtgccagccc
     4081 tgggaccgaa ccccgcgttt atgaacaaac gacccaacac cgtgcgtttt attctgtctt
     4141 tttattgccg tcatagcgcg ggttccttcc ggtattgtct ccttccgtgt ttcagttagc
     4201 ctccccctag ggtgggcgaa gaactccagc atgagatccc cgcgctggag gatcatccag
     4261 ccggcgtccc ggaaaacgat tccgaagccc aacctttcat agaaggcggc ggtggaatcg
     4321 aaatctcgtg atggcaggtt gggcgtcgct tggtcggtca tttcgaaccc cagagtcccg
     4381 ctcagaagaa ctcgtcaaga aggcgataga aggcgatgcg ctgcgaatcg ggagcggcga
     4441 taccgtaaag cacgaggaag cggtcagccc attcgccgcc aagctcttca gcaatatcac
     4501 gggtagccaa cgctatgtcc tgatagcggt ccgccacacc cagccggcca cagtcgatga
     4561 atccagaaaa gcggccattt tccaccatga tattcggcaa gcaggcatcg ccatgggtca
     4621 cgacgagatc ctcgccgtcg ggcatgctcg ccttgagcct ggcgaacagt tcggctggcg
     4681 cgagcccctg atgctcttcg tccagatcat cctgatcgac aagaccggct tccatccgag
     4741 tacgtgctcg ctcgatgcga tgtttcgctt ggtggtcgaa tgggcaggta gccggatcaa
     4801 gcgtatgcag ccgccgcatt gcatcagcca tgatggatac tttctcggca ggagcaaggt
     4861 gagatgacag gagatcctgc cccggcactt cgcccaatag cagccagtcc cttcccgctt
     4921 cagtgacaac gtcgagcaca gctgcgcaag gaacgcccgt cgtggccagc cacgatagcc
     4981 gcgctgcctc gtcttgcagt tcattcaggg caccggacag gtcggtcttg acaaaaagaa
     5041 ccgggcgccc ctgcgctgac agccggaaca cggcggcatc agagcagccg attgtctgtt
     5101 gtgcccagtc atagccgaat agcctctcca cccaagcggc cggagaacct gcgtgcaatc
     5161 catcttgttc aatcatgcga aacgatcctc atcctgtctc ttgatcgatc tttgcaaaag
     5221 cctaggcctc caaaaaagcc tcctcactac ttctggaata gctcagaggc cgaggcggcc
     5281 tcggcctctg cataaataaa aaaaattagt cagccatggg gcggagaatg ggcggaactg
     5341 ggcggagtta ggggcgg
//
