LOCUS       Exported                6374 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    Cortactin-pmCherryC1
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6374)
  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 6374)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6374
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     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    3102..3223
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3139..3158)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3193..3212
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3230..3685)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3367..3388)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3579..3598
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3712..3816
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3818..4175
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     primer_bind     complement(3841..3861)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4026..4161
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4088..4107
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4210..5004
                     /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(4264..4283)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4874..4893
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5192..5211)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5236..5283
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5612..6200
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6101..6120
                     /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 ctcgagctca agcttcgaat tctgcagtcg acggtaccat gtggaaagcc tctgcaggcc
     1321 atgctgtgtc catcacgcag gatgatggag gagctgatga ctgggagact gatcctgatt
     1381 ttgtgaatga tgtgagtgaa aaggagcaga gatggggtgc taaaaccgtg cagggatcgg
     1441 ggcaccagga acacatcaac attcacaagc ttcgagagaa tgtcttccaa gaacaccaga
     1501 cgctcaagga gaaggagctg gaaacgggac ccaaggcttc ccacggctat ggcgggaagt
     1561 tcggtgtgga gcaggatagg atggacagat cagccgtggg ccatgagtac cagtcgaagc
     1621 tttccaagca ctgctcacaa gtggactcgg tccggggctt cggaggcaag ttcggtgtcc
     1681 agatggacag ggtggatcag tctgctgtag gctttgaata ccaggggaag actgagaagc
     1741 atgcctccca gaaagactac tctagtggct tcggtggcaa atacggtgtg caagctgacc
     1801 gtgtagacaa gagtgccgtg ggctttgact accagggcaa gacggagaag catgagtctc
     1861 agaaagatta ctccaaaggt tttggtggca aatatgggat tgacaaggac aaggtggata
     1921 aaagtgctgt gggctttgag tatcaaggca agacagagaa gcacgaatcc cagaaagact
     1981 atgtaaaagg ctttggagga aagtttggtg tgcagacaga cagacaggac aagtgtgccc
     2041 ttggctggga ccatcaggag aagctgcagc tgcatgaatc ccaaaaagac tataagactg
     2101 gtttcggagg caaatttggt gttcagtccg agaggcagga ctcctccgct gtggggtttg
     2161 attacaagga gagattggcc aagcacgagc cccagcaaga ctatgccaaa ggattcggcg
     2221 ggaagtatgg ggtgcagaag gatcggatgg acaagaatgc atccaccttt gaagaagtgg
     2281 tccaggtgcc atctgcctat cagaagactg tccccattga ggccgtaacc agcaaaacca
     2341 gtaatatccg tgctaacttt gaaaacctgg caaaggagag agagcaggag gacaggcgga
     2401 aggcagaagc cgagagagct cagcggatgg ccaaagaaag acaggagcag gaggaggcgc
     2461 gcaggaagct ggaagagcaa gccagagcca agaagcagac gccccctgca tcccctagtc
     2521 ctcaaccaat tgaagacaga ccaccctcca gccccatcta tgaggatgca gctccgttca
     2581 aggccgagcc gagctaccga ggtagcgaac ctgagcctga gtacagcatc gaggccgcag
     2641 gcattcctga ggctggcagc cagcaaggcc tgacctatac atcagagccc gtgtacgaga
     2701 ctacagaggc tcctggccac tatcaagcag aggatgacac ctacgatggg tatgagagtg
     2761 acctgggcat cacagccatc gccctgtatg actaccaggc tgctggcgat gatgagatct
     2821 cctttgaccc tgatgacatc atcaccaaca tagaaatgat tgacgatggc tggtggcgtg
     2881 gggtgtgcaa gggcagatac gggctcttcc cagccaacta tgtggagctg cggcagtagg
     2941 aattctgcag atatccatca cactggcggc cgctcgagca tgcatctaga taactgatca
     3001 taatcagcca taccacattt gtagaggttt tacttgcttt aaaaaacctc ccacacctcc
     3061 ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt taacttgttt attgcagctt
     3121 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     3181 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttaacgcgta aattgtaagc
     3241 gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa
     3301 taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt
     3361 gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg
     3421 cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta atcaagtttt
     3481 ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga
     3541 gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg
     3601 ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg
     3661 cttaatgcgc cgctacaggg cgcgtcaggt ggcacttttc ggggaaatgt gcgcggaacc
     3721 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     3781 tgataaatgc ttcaataata ttgaaaaagg aagagtcctg aggcggaaag aaccagctgt
     3841 ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     3901 aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag
     3961 gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc
     4021 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
     4081 ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt
     4141 gaggaggctt ttttggaggc ctaggctttt gcaaagatcg atcaagagac aggatgagga
     4201 tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag
     4261 aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc
     4321 cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg
     4381 aatgaactgc aagacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc
     4441 gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg
     4501 ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct
     4561 gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg
     4621 aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat
     4681 ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgagc
     4741 atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg
     4801 gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc
     4861 tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct
     4921 gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat
     4981 cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga
     5041 cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct
     5101 tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg
     5161 agttcttcgc ccaccctagg gggaggctaa ctgaaacacg gaaggagaca ataccggaag
     5221 gaacccgcgc tatgacggca ataaaaagac agaataaaac gcacggtgtt gggtcgtttg
     5281 ttcataaacg cggggttcgg tcccagggct ggcactctgt cgatacccca ccgagacccc
     5341 attggggcca atacgcccgc gtttcttcct tttccccacc ccacccccca agttcgggtg
     5401 aaggcccagg gctcgcagcc aacgtcgggg cggcaggccc tgccatagcc tcaggttact
     5461 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     5521 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     5581 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     5641 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     5701 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     5761 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     5821 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     5881 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     5941 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     6001 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     6061 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     6121 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     6181 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     6241 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     6301 ttaccgccat gcattagtta ttaatagtaa tcaattacgg ggtcattagt tcatagccca
     6361 tatatggagt tccg
//
