LOCUS       Exported                7347 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    Dyn2-pmCherryN1
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7347)
  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 7347)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7347
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        1..204
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     155..175
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      2933..2942
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             2939..3643
                     /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(3084..3102)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3352..3371
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    3771..3892
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3808..3827)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3862..3881
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3899..4354)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4036..4057)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4248..4267
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4381..4485
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4487..4844
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     primer_bind     complement(4510..4530)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4695..4830
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4757..4776
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4879..5673
                     /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(4933..4952)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5543..5562
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5861..5880)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5905..5952
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6281..6869
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6770..6789
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        7044..7347
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
       61 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
      121 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
      181 tgggaggtct atataagcag agctggttta gtgaaccgtc agatccgcta gcgctaccgg
      241 actcagatct accatgggca accgcgggat ggaagagctg atcccgctgg tcaacaaact
      301 gcaggacgcc ttcagctcca tcggccagag ctgccacctg gacctgccgc agatcgctgt
      361 agtgggcggc cagagcgccg gcaagagctc ggtgctggag aacttcgtgg gccgggactt
      421 ccttccccgc ggttcaggaa tcgtcacccg gcggcctctc attctgcagc tcatcttctc
      481 aaaaacagaa catgccgagt ttttgcactg caagtccaaa aagtttacag actttgatga
      541 agtccggcag gagattgaag cagagaccga cagggtcacg gggaccaaca aaggcatctc
      601 cccagtgccc atcaaccttc gagtctactc gccacacgtg ttgaacttga ccctcatcga
      661 cctcccgggt atcaccaagg tgcctgtggg cgaccagcct ccagacatcg agtaccagat
      721 caaggacatg atcctgcagt tcatcagccg ggagagcagc ctcattctgg ctgtcacgcc
      781 cgccaacatg gacctggcca actccgacgc cctcaagctg gccaaggaag tcgatcccca
      841 aggcctacgg accatcggtg tcatcaccaa gcttgacctg atggacgagg gcaccgacgc
      901 cagggacgtc ttggagaaca agttgctccc gttgagaaga ggctacattg gcgtggtgaa
      961 ccgcagccag aaggatattg agggcaagaa ggacatccgt gcagcactgg cagctgagag
     1021 gaagttcttc ctctcccacc cggcctaccg gcacatggcc gaccgcatgg gcacgccaca
     1081 tctgcagaag acgctgaatc agcaactgac caaccacatc cgggagtcgc tgccggccct
     1141 acgtagcaaa ctacagagcc agctgctgtc cctggagaag gaggtggagg agtacaagaa
     1201 ctttcggccc gacgacccca cccgcaaaac caaagccctg ctgcagatgg tccagcagtt
     1261 tggggtggat tttgagaaga ggatcgaggg ctcaggagat caggtggaca ctctggagct
     1321 ctccgggggc gcccgaatca atcgcatctt ccacgagcgg ttcccatttg agctggtgaa
     1381 gatggagttt gacgagaagg acttacgacg ggagatcagc tatgccatta agaacatcca
     1441 tggagtcagg accgggcttt tcaccccgga cttggcattc gaggccattg tgaaaaagca
     1501 ggtcgtcaag ctgaaagagc cctgtctgaa atgtgtcgac ctggttatcc aggagctaat
     1561 caatacagtt aggcagtgta ccagtaagct cagttcctac ccccggttgc gagaggagac
     1621 agagcgaatc gtcaccactt acatccggga acgggagggg agaacgaagg accagattct
     1681 tctgctgatc gacattgagc agtcctacat caacacgaac catgaggact tcatcgggtt
     1741 tgccaatgcc cagcagagga gcacgcagct gaacaagaag agagccatcc ccaatcaggt
     1801 gatccgcagg ggctggctga ccatcaacaa catcagcctg atgaaaggcg gctccaagga
     1861 gtactggttt gtgctgactg ccgagtcact gtcctggtac aaggatgagg aggagaaaga
     1921 gaagaagtac atgctgcctc tggacaacct caagatccgt gatgtggaga agggcttcat
     1981 gtccaacaag cacgtcttcg ccatcttcaa cacggagcag agaaacgtct acaaggacct
     2041 gcggcagatc gagctggcct gtgactccca ggaagacgtg gacagctgga aggcctcgtt
     2101 cctccgagct ggcgtctacc ccgagaagga ccaggcagaa aacgaggatg gggcccagga
     2161 gaacaccttc tccatggacc cccaactgga gcggcaggtg gagaccattc gcaacctggt
     2221 ggactcatac gtggccatca tcaacaagtc catccgcgac ctcatgccaa agaccatcat
     2281 gcacctcatg atcaacaata cgaaggcctt catccaccac gagctgctgg cctacctata
     2341 ctcctcggca gaccagagca gcctcatgga ggagtcggct gaccaggcac agcggcggga
     2401 cgacatgctg cgcatgtacc atgccctcaa ggaggcgctc aacatcatcg gtgacatcag
     2461 caccagcact gtgtccacgc ctgtaccccc gcctgtcgat gacacctggc tccagagcgc
     2521 cagcagccac agccccactc cacagcgccg accggtgtcc agcatacacc cccctggccg
     2581 gcccccagca gtgaggggcc ccactccagg gccccccctg attcctgttc ccgtgggggc
     2641 agcagcctcc ttctcggcgc ccccaatccc atcccggcct ggaccccaga gcgtgtttgc
     2701 caacagtgac ctcttcccag ccccgcctca gatcccatct cggccagttc ggatcccccc
     2761 agggattccc ccaggagtgc ccagcagaag accccctgct gcgcccagcc ggcccaccat
     2821 tatccgccca gccgagccat ccctgctcga cggacatggt acaggatcaa ctggtagtgg
     2881 aagtagtgga attctgcagt cgacggtacc gcgggcccgg gatccaccgg tcgccaccat
     2941 ggtgagcaag ggcgaggagg ataacatggc catcatcaag gagttcatgc gcttcaaggt
     3001 gcacatggag ggctccgtga acggccacga gttcgagatc gagggcgagg gcgagggccg
     3061 cccctacgag ggcacccaga ccgccaagct gaaggtgacc aagggtggcc ccctgccctt
     3121 cgcctgggac atcctgtccc ctcagttcat gtacggctcc aaggcctacg tgaagcaccc
     3181 cgccgacatc cccgactact tgaagctgtc cttccccgag ggcttcaagt gggagcgcgt
     3241 gatgaacttc gaggacggcg gcgtggtgac cgtgacccag gactcctccc tgcaggacgg
     3301 cgagttcatc tacaaggtga agctgcgcgg caccaacttc ccctccgacg gccccgtaat
     3361 gcagaagaag accatgggct gggaggcctc ctccgagcgg atgtaccccg aggacggcgc
     3421 cctgaagggc gagatcaagc agaggctgaa gctgaaggac ggcggccact acgacgctga
     3481 ggtcaagacc acctacaagg ccaagaagcc cgtgcagctg cccggcgcct acaacgtcaa
     3541 catcaagttg gacatcacct cccacaacga ggactacacc atcgtggaac agtacgaacg
     3601 cgccgagggc cgccactcca ccggcggcat ggacgagctg tacaataaag cggccgcgac
     3661 tctagatcat aatcagccat accacatttg tagaggtttt acttgcttta aaaaacctcc
     3721 cacacctccc cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta
     3781 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     3841 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct taaggcgtaa
     3901 attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt
     3961 tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata
     4021 gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac
     4081 gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa
     4141 tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc
     4201 cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg
     4261 aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca
     4321 cccgccgcgc ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg
     4381 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     4441 caataaccct gataaatgct tcaataatat tgaaaaagga agagtcctga ggcggaaaga
     4501 accagctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca
     4561 gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct
     4621 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc
     4681 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
     4741 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc
     4801 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaagatcga tcaagagaca
     4861 ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct
     4921 tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc
     4981 gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc
     5041 ggtgccctga atgaactgca agacgaggca gcgcggctat cgtggctggc cacgacgggc
     5101 gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg
     5161 ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc
     5221 atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac
     5281 caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat
     5341 caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc
     5401 aaggcgagca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg
     5461 aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg
     5521 gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc
     5581 gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc
     5641 gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg
     5701 accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa
     5761 ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc
     5821 tcatgctgga gttcttcgcc caccctaggg ggaggctaac tgaaacacgg aaggagacaa
     5881 taccggaagg aacccgcgct atgacggcaa taaaaagaca gaataaaacg cacggtgttg
     5941 ggtcgtttgt tcataaacgc ggggttcggt cccagggctg gcactctgtc gataccccac
     6001 cgagacccca ttggggccaa tacgcccgcg tttcttcctt ttccccaccc caccccccaa
     6061 gttcgggtga aggcccaggg ctcgcagcca acgtcggggc ggcaggccct gccatagcct
     6121 caggttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     6181 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     6241 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     6301 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     6361 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     6421 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     6481 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     6541 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     6601 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     6661 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     6721 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     6781 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     6841 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     6901 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
     6961 ataaccgtat taccgccatg cattagttat taatagtaat caattacggg gtcattagtt
     7021 catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga
     7081 ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca
     7141 atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca
     7201 gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg
     7261 cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc
     7321 tacgtattag tcatcgctat taccatg
//
