LOCUS       sequence-338153-        7088 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 7088)
  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 7088)
  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: FCHo2-pmCherryC1
FEATURES             Location/Qualifiers
     source          1..7088
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             540..1247
                     /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(685..703)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     953..972
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    3816..3937
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3853..3872)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3907..3926
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3944..4399)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4081..4102)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4293..4312
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4426..4530
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4532..4889
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4532..4723
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4555..4575)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4740..4875
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4924..5718
                     /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             4924..5718
                     /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(4978..4997)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5588..5607
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5906..5925)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5950..5997
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6326..6914
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6815..6834
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcacca
      541 tggtgagcaa gggcgaggag gataacatgg ccatcatcaa ggagttcatg cgcttcaagg
      601 tgcacatgga gggctccgtg aacggccacg agttcgagat cgagggcgag ggcgagggcc
      661 gcccctacga gggcacccag accgccaagc tgaaggtgac caagggtggc cccctgccct
      721 tcgcctggga catcctgtcc cctcagttca tgtacggctc caaggcctac gtgaagcacc
      781 ccgccgacat ccccgactac ttgaagctgt ccttccccga gggcttcaag tgggagcgcg
      841 tgatgaactt cgaggacggc ggcgtggtga ccgtgaccca ggactcctcc ctgcaggacg
      901 gcgagttcat ctacaaggtg aagctgcgcg gcaccaactt cccctccgac ggccccgtaa
      961 tgcagaagaa gaccatgggc tgggaggcct cctccgagcg gatgtacccc gaggacggcg
     1021 ccctgaaggg cgagatcaag cagaggctga agctgaagga cggcggccac tacgacgctg
     1081 aggtcaagac cacctacaag gccaagaagc ccgtgcagct gcccggcgcc tacaacgtca
     1141 acatcaagtt ggacatcacc tcccacaacg aggactacac catcgtggaa cagtacgaac
     1201 gcgccgaggg ccgccactcc accggcggca tggacgagct gtacaagtcc ggactcagat
     1261 ctcgagctcg gatgggttcg acctcgggtt cgaccaaggg tcagctgatg aagcatggac
     1321 agatatcaac taaagaacta gcagattttg taagagaaag agctacaata gaagaggcat
     1381 actccagatc aatgacaaaa ctagcgaaat ctgcaagcaa ttattcacaa cttggaacat
     1441 ttgcaccaat gtgggatgta ttcaaaacat ctacagagaa attagcaaat tgtcatttgg
     1501 atcttgttag aaaattacag gaattaataa aggaagttca gaagtatgga gaggagcaag
     1561 tcaagtctca taagaagact aaagaagaag tcgcagggac tctggaagca gtccaagcca
     1621 ttcagaatat aactcaggca ctccagaagt cgaaggaaaa ttacactgcc aaatgtgtag
     1681 aacaggaacg tctgaaaaag gaaggagcta cacaaagaga aatagaaaag gcagctgtta
     1741 agtctaagaa agctacagat acttataaac tttatgtgga aaagtatgca ttaacaaaag
     1801 cagattttga gcagaagatg acagaaacag cccagaaatt tcaagatatt gaagaaactc
     1861 acctcattca cataaaggaa attatcggat ccttgtcaaa tgctgtcaag gagatccact
     1921 tacagatcgg ccaggttcat gaagagttta taaataacat ggctaatact acaattgaaa
     1981 gtttgattca gaaatttgcc gagtcaaaag gcacagggaa ggaaagacct ggtcttattg
     2041 aatttgaaga gtgtgaccct gctagtgcag ttgaaggcat aaaaccaagg aaaagaaaga
     2101 cttttgcttt gcctggaata attaaaaagg aaaaggacgc agaatctgtg gaatgtcctg
     2161 atgcagattc gcttaatatt ccagatgtag atgaagaagg tttcagtatt aaaccagaag
     2221 caaatcagaa tgataccaaa gagaaccatt tctactcatc cagtgactct gactctgaag
     2281 atgaagaacc aaagaggtac cggatagaga tcaagcccgc gcatccaaat aacttgcatc
     2341 acacaatggc ttctttggat gaactgaaag tatccatagg gaatataaca ttgtccccag
     2401 cagtatctag acacagtcca gtgcagatga atcggaattc atctaatgaa gaattaacaa
     2461 aatcaaagcc gtcttctcta cccactgaga aagggaccaa tgatttactt gcttgggatc
     2521 ccctgttcgg atcgtcgctt gagtcatctt cagctccact aacttcgtca tcatcggcca
     2581 gacccacgac acctctctct ttaggcaccc ttgttccacc tccaaggcct gcttccagac
     2641 caaagcttgc ttcaggaaaa ctgagtggga tcaatgaaat acccaggcca ttcagcccac
     2701 ctgtaacttc caacaccagc ccacctccta ctgctccctt agcccgagcc gagagcagct
     2761 cctctatctc ctcgtctgcc tcgttgagtg ctgccaacac tccaacagta ggtgtatcac
     2821 ggggtcccag cccggtcagc cttggaaacc aggacacttt acctgtggca atcgccctta
     2881 cagaatctgt caacgcctac tttaaaggag cagatcctac caagtgtatt gtgaagatca
     2941 cgggagacgt gacaatatca tttcccagtg ggattattaa agtcttcacc agcaacccct
     3001 ccccagctgt gctgtgcttt agggtgaaga acatcagcag gctggagcag atccttccaa
     3061 actcccagct tgtgttcagt gatccatcac agtgtgattc caacacaaaa gatttttgga
     3121 tgaacatgca agctgttact atctacctca agaagctatc agaacaaaat ccagctgctt
     3181 cttattataa tgtagatgta ttaaaatacc aggtatcatc aaatggcatt cagtccactc
     3241 ctctaaatct tgcaacctac tggaaatgta gtgccagcac cacagatctg agagtggatt
     3301 ataagtacaa cccagaagct atggtcgctc cgagtgtgct ttccaacata caggtggtgg
     3361 ttcccgtgga tggaggtgtc acaaatatgc agtcccttcc tcctgcaata tggaatgcag
     3421 agcaaatgaa agctttttgg aaattatctg ggatttcgga gaaatcagac agtggaggtt
     3481 ctggatccct cagagcaaaa tttgatcttt cagaaggacc cagcaaacct acaacactgg
     3541 cagtgcaatt cctcagtgag ggcaatacgc tctcaggagt ggatattgag cttgtgggta
     3601 ctggctatag actttcctta gtaaagaagc gctttgccac tggacgatac ttggcagatt
     3661 gttgagtcga cggtaccgcg ggcccgggat ccaccggatc tagataactg atcataatca
     3721 gccataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac ctccccctga
     3781 acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca gcttataatg
     3841 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     3901 ctagttgtgg tttgtccaaa ctcatcaatg tatcttaacg cgtaaattgt aagcgttaat
     3961 attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc
     4021 gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt
     4081 ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa
     4141 accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg
     4201 tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga
     4261 cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct
     4321 agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat
     4381 gcgccgctac agggcgcgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     4441 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     4501 atgcttcaat aatattgaaa aaggaagagt cctgaggcgg aaagaaccag ctgtggaatg
     4561 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     4621 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa
     4681 gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca
     4741 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
     4801 ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag
     4861 gcttttttgg aggcctaggc ttttgcaaag atcgatcaag agacaggatg aggatcgttt
     4921 cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta
     4981 ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg
     5041 tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa
     5101 ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct
     5161 gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg
     5221 caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca
     5281 atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat
     5341 cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac
     5401 gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gagcatgccc
     5461 gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa
     5521 aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag
     5581 gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc
     5641 ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt
     5701 cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca
     5761 acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa
     5821 tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct
     5881 tcgcccaccc tagggggagg ctaactgaaa cacggaagga gacaataccg gaaggaaccc
     5941 gcgctatgac ggcaataaaa agacagaata aaacgcacgg tgttgggtcg tttgttcata
     6001 aacgcggggt tcggtcccag ggctggcact ctgtcgatac cccaccgaga ccccattggg
     6061 gccaatacgc ccgcgtttct tccttttccc caccccaccc cccaagttcg ggtgaaggcc
     6121 cagggctcgc agccaacgtc ggggcggcag gccctgccat agcctcaggt tactcatata
     6181 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     6241 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     6301 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     6361 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     6421 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     6481 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     6541 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     6601 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     6661 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     6721 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     6781 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     6841 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     6901 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     6961 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     7021 ccatgcatta gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg
     7081 gagttccg
//
