LOCUS       sequence-396876-        6497 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 6497)
  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 6497)
  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: Epsin2-pmCherryC1
FEATURES             Location/Qualifiers
     source          1..6497
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        117..420
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        421..624
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     575..595
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             656..1363
                     /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(801..819)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1069..1088
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    3341..3462
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3378..3397)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3432..3451
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3469..3924)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3606..3627)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3818..3837
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3951..4055
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4057..4414
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4057..4248
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4080..4100)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4265..4400
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4449..5243
                     /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             4449..5243
                     /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(4503..4522)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5113..5132
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5431..5450)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5475..5522
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5851..6439
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6340..6359
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc atgcattagt
       61 tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt
      121 acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg
      181 tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg
      241 gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt
      301 acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg
      361 accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg
      421 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
      481 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
      541 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
      601 tgggaggtct atataagcag agctggttta gtgaaccgtc agatccgcta gcaccatggt
      661 gagcaagggc gaggaggata acatggccat catcaaggag ttcatgcgct tcaaggtgca
      721 catggagggc tccgtgaacg gccacgagtt cgagatcgag ggcgagggcg agggccgccc
      781 ctacgagggc acccagaccg ccaagctgaa ggtgaccaag ggtggccccc tgcccttcgc
      841 ctgggacatc ctgtcccctc agttcatgta cggctccaag gcctacgtga agcaccccgc
      901 cgacatcccc gactacttga agctgtcctt ccccgagggc ttcaagtggg agcgcgtgat
      961 gaacttcgag gacggcggcg tggtgaccgt gacccaggac tcctccctgc aggacggcga
     1021 gttcatctac aaggtgaagc tgcgcggcac caacttcccc tccgacggcc ccgtaatgca
     1081 gaagaagacc atgggctggg aggcctcctc cgagcggatg taccccgagg acggcgccct
     1141 gaagggcgag atcaagcaga ggctgaagct gaaggacggc ggccactacg acgctgaggt
     1201 caagaccacc tacaaggcca agaagcccgt gcagctgccc ggcgcctaca acgtcaacat
     1261 caagttggac atcacctccc acaacgagga ctacaccatc gtggaacagt acgaacgcgc
     1321 cgagggccgc cactccaccg gcggcatgga cgagctgtac aagtccggac tcagatctcg
     1381 agctcggatg ggttcgacct cgggttcgac caagggtcag ctgatgacaa cttcatctat
     1441 cagacggcag atgaaaaaca ttgtgaacaa ttactcagag gctgaaatca aagtccggga
     1501 ggccacctcc aatgacccgt ggggcccatc cagctctctg atgactgaga ttgctgacct
     1561 gacctacaat gtggtagcgt tctcggagat catgagcatg gtttggaagc gacttaatga
     1621 ccatggcaag aactggcggc atgtatacaa ggcactgaca ctgctggact accttatcaa
     1681 gacaggttct gagcgggtgg cccagcagtg ccgtgagaac atctttgcca tacagactct
     1741 gaaggacttt cagtacattg accgtgatgg caaagaccag ggtattaacg ttcgagagaa
     1801 atcaaagcaa ctggttgcgc tcctcaagga tgaggagcgg ctgaaggttg agagagttca
     1861 ggctctcaaa accaaagagc gcatggctca ggtggccact ggcgtgggca gcaaccaaat
     1921 cacctttggc cgtggctcca gccagcccaa cctttctacc agctactcag agcaggagta
     1981 tggcaaggct gggggctcgc cggcatccta ccacggctct acttccccac gagtatcctc
     2041 tgagttagag caggcccggc cacagacaag cggagaagag gagctgcagc tacaactggc
     2101 acttgccatg agcagagagg ttgctgagca gtcatcagaa agcgtgcaga cagccagagg
     2161 cagcaaggaa gaacgcctca ggcggggtga tgacctcaga ttgcagatgg ctctggaaga
     2221 aagccggaga gacacagtaa aagttccaaa aaagaaagag gcgaaagctt gttgcaagcc
     2281 aggctcccac tcccagcaga ctaccttgtt ggatttaatg gatgccctcc ccagctcagg
     2341 ccctgttaca cagaaaactg agccctggag tgcaggagcc tctgctaacc agaccaaccc
     2401 ctggggtgga acagtggctc cttcgaacat tactgacccg tggccatcgt ttggtaccaa
     2461 gccagctgcc tctgtggacc cctggggagt acctaccaca gccagcacac agtctgtccc
     2521 taagaactca gacccttggg cagcctcaca gcagcctgcc tccaatgctg gaaagacaac
     2581 agatgcctgg ggagctgcta agcccagttc tgcctcaggg tcctttgagc tcttcagtaa
     2641 tttcaacggt acagttaaag atgatttttc tgaatttgac aaccttcgaa cttcaaaaaa
     2701 accagctgag tcaggggcct cagtgccacc ccaggacagc agaaccacga gccctgacct
     2761 ctttgagtct caatccttga cttctgcgtc aagcaagcct agcagtgcca ggaaaacacc
     2821 tgagtccttc ctgggcccca atgcagccct ggtgaacctg gactcattgg tgactaagcc
     2881 tgctccacca gctcagtccc ttaatccctt cctggcacca ggtgctgctg ctccagcccc
     2941 tgtcaacccc ttccaggtca accagcccca gccgctgaca ctgaaccagc tccggggaag
     3001 ccctgtcctg ggaagcagtg catcctttgg gtctggtcca ggggtggaga ctgtggctcc
     3061 catgacctca gtagctccac actcatcagt gggggccagt ggctcctcct tgacaccact
     3121 gggccctaca gcaatgaaca tggtaggcag tgtgggcatt cccccatcag cagctcagtc
     3181 aacaggcaca acaaaccctt tccttctcta gggatccacc ggatctagat aactgatcat
     3241 aatcagccat accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc
     3301 cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta
     3361 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     3421 gcattctagt tgtggtttgt ccaaactcat caatgtatct taacgcgtaa attgtaagcg
     3481 ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat
     3541 aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg
     3601 ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc
     3661 gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt
     3721 tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag
     3781 cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg
     3841 gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc
     3901 ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc
     3961 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     4021 gataaatgct tcaataatat tgaaaaagga agagtcctga ggcggaaaga accagctgtg
     4081 gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     4141 aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg
     4201 cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc
     4261 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
     4321 tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg
     4381 aggaggcttt tttggaggcc taggcttttg caaagatcga tcaagagaca ggatgaggat
     4441 cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga
     4501 ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc
     4561 ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga
     4621 atgaactgca agacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg
     4681 cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc
     4741 cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg
     4801 atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga
     4861 aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc
     4921 tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgagca
     4981 tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg
     5041 tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct
     5101 atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg
     5161 accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc
     5221 gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac
     5281 gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt
     5341 cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga
     5401 gttcttcgcc caccctaggg ggaggctaac tgaaacacgg aaggagacaa taccggaagg
     5461 aacccgcgct atgacggcaa taaaaagaca gaataaaacg cacggtgttg ggtcgtttgt
     5521 tcataaacgc ggggttcggt cccagggctg gcactctgtc gataccccac cgagacccca
     5581 ttggggccaa tacgcccgcg tttcttcctt ttccccaccc caccccccaa gttcgggtga
     5641 aggcccaggg ctcgcagcca acgtcggggc ggcaggccct gccatagcct caggttactc
     5701 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     5761 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     5821 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     5881 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     5941 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     6001 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     6061 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     6121 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     6181 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     6241 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     6301 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     6361 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     6421 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     6481 ctggcctttt gctcaca
//
