LOCUS       Exported                7014 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pTfR-PAmKate
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7014)
  AUTHORS   Gunewardene MS, Subach FV, Gould TJ, Penoncello GP, Gudheti MV, 
            Verkhusha VV, Hess ST
  TITLE     Superresolution imaging of multiple fluorescent proteins with highly
            overlapping emission spectra in living cells.
  JOURNAL   Biophys J. 2011 Sep 21;101(6):1522-8. Epub 2011 Sep 20.
  PUBMED    21943434
REFERENCE   2  (bases 1 to 7014)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7014
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             complement(1565..1576)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     regulatory      2960..2969
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             2966..3679
                     /codon_start=1
                     /product="monomeric photoactivatable red fluorescent 
                     protein (Gunewardene et al., 2011)"
                     /label=PAmKate
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTM
                     RIKVVEGGPLPFAFDILATSFMYGSKTFINHTQGIPDFWKQSFPEGFTWERVTTYEDGG
                     VLTATQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEANTEMLYPADGGLEGRGD
                     MALKLVGGGHLICNLKTTYRSKKPAKNLKMPGVYYVDRRLERIKEADKETYVEQHEVAV
                     ARYCDLPSKLGHKLN"
     polyA_signal    3802..3923
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3839..3858)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3893..3912
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3930..4385)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4067..4088)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4279..4298
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4412..4516
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4518..4875
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     primer_bind     complement(4541..4561)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4726..4861
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4788..4807
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4910..5704
                     /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(4964..4983)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5574..5593
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5892..5911)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5936..5983
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6312..6900
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6801..6820
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggactcag atctcgagcg ccaccatgat ggatcaagct agatcagcat tctctaactt
      661 gtttggtgga gaaccattgt catatacccg gttcagcctg gctcggcaag tagatggcga
      721 taacagtcat gtggagatga aacttgctgt agatgaagaa gaaaatgctg acaataacac
      781 aaaggccaat gtcacaaaac caaaaaggtg tagtggaagt atctgctatg ggactattgc
      841 tgtgatcgtc tttttcttga ttggatttat gattggctac ttgggctatt gtaaaggggt
      901 agaaccaaaa actgagtgtg agagactggc aggaaccgag tctccagtga gggaggagcc
      961 aggagaggac ttccctgcag cacgtcgctt atattgggat gacctgaaga gaaagttgtc
     1021 ggagaaactg gacagcacag acttcaccag caccatcaag ctgctgaatg aaaattcata
     1081 tgtccctcgt gaggctggat ctcaaaaaga tgaaaatctt gcgttgtatg ttgaaaatca
     1141 atttcgtgaa tttaaactca gcaaagtctg gcgtgatcaa cattttgtta agattcaggt
     1201 caaagacagc gctcaaaact cggtgatcat agttgataag aacggtagac ttgtttacct
     1261 ggtggagaat cctgggggtt atgtggcgta tagtaaggct gcaacagtta ctggtaaact
     1321 ggtccatgct aattttggta ctaaaaaaga ttttgaggat ttatacactc ctgtgaatgg
     1381 atctatagtg attgtcagag cagggaaaat cacctttgca gaaaaggttg caaatgctga
     1441 aagcttaaat gcaattggtg tgttgatata catggaccag actaaatttc ccattgttaa
     1501 cgcagaactt tcattctttg gacatgctca tctggggaca ggtgaccctt acacacctgg
     1561 attcccttcc ttcaatcaca ctcagtttcc accatctcgg tcatcaggat tgcctaatat
     1621 acctgtccag acaatctcca gagctgctgc agaaaagctg tttgggaata tggaaggaga
     1681 ctgtccctct gactggaaaa cagactctac atgtaggatg gtaacctcag aaagcaagaa
     1741 tgtgaagctc actgtgagca atgtgctgaa agagataaaa attcttaaca tctttggagt
     1801 tattaaaggc tttgtagaac cagatcacta tgttgtagtt ggggcccaga gagatgcatg
     1861 gggccctgga gctgcaaaat ccggtgtagg cacagctctc ctattgaaac ttgcccagat
     1921 gttctcagat atggtcttaa aagatgggtt tcagcccagc agaagcatta tctttgccag
     1981 ttggagtgct ggagactttg gatcggttgg tgccactgaa tggctagagg gatacctttc
     2041 gtccctgcat ttaaaggctt tcacttatat taatctggat aaagcggttc ttggtaccag
     2101 caacttcaag gtttctgcca gcccactgtt gtatacgctt attgagaaaa caatgcaaaa
     2161 tgtgaagcat ccggttactg ggcaatttct atatcaggac agcaactggg ccagcaaagt
     2221 tgagaaactc actttagaca atgctgcttt ccctttcctt gcatattctg gaatcccagc
     2281 agtttctttc tgtttttgcg aggacacaga ttatccttat ttgggtacca ccatggacac
     2341 ctataaggaa ctgattgaga ggattcctga gttgaacaaa gtggcacgag cagctgcaga
     2401 ggtcgctggt cagttcgtga ttaaactaac ccatgatgtt gaattgaacc tggactatga
     2461 gaggtacaac agccaactgc tttcatttgt gagggatctg aaccaataca gagcagacat
     2521 aaaggaaatg ggcctgagtt tacagtggct gtattctgct cgtggagact tcttccgtgc
     2581 tacttccaga ctaacaacag atttcgggaa tgctgagaaa acagacagat ttgtcatgaa
     2641 gaaactcaat gatcgtgtca tgagagtgga gtatcacttc ctctctccct acgtatctcc
     2701 aaaagagtct cctttccgac atgtcttctg gggctccggc tctcacacgc tgccagcttt
     2761 actggagaac ttgaaactgc gtaaacaaaa taacggtgct tttaatgaaa cgctgttcag
     2821 aaaccagttg gctctagcta cttggactat tcagggagct gcaaatgccc tctctggtga
     2881 cgtttgggac attgacaatg agttttcaga attcggctcc accggctcca ccggctccac
     2941 cggcgcggat ccaccggtcg ccaccatggt gtctaagggc gaagagctga ttaaggagaa
     3001 catgcacatg aagctgtaca tggagggcac cgtgaacaac caccacttca agtgcacatc
     3061 cgagggcgaa ggcaagccct acgagggcac ccagaccatg agaatcaagg tggtcgaggg
     3121 cggccctctc cccttcgcct tcgacatcct ggctaccagc ttcatgtacg gcagcaagac
     3181 cttcatcaac cacacccagg gcatccccga cttctggaag cagtccttcc ctgagggctt
     3241 cacatgggag agagtcacca catacgaaga cgggggcgtg ctgaccgcta cccaggacac
     3301 cagcctccag gacggctgcc tcatctacaa cgtcaagatc agaggggtga acttcccatc
     3361 caacggccct gtgatgcaga agaaaacact cggctgggag gccaacaccg agatgctgta
     3421 ccccgctgac ggcggcctgg aaggcagagg ggacatggcc ctgaagctcg tgggcggggg
     3481 ccacctgatc tgcaacttga agaccacata cagatccaag aaacccgcta agaacctcaa
     3541 gatgcccggc gtctactatg tggaccgcag actggaaaga atcaaggagg ccgacaaaga
     3601 gacctacgtc gagcagcacg aggtggctgt ggccagatac tgcgacctcc ctagcaaact
     3661 ggggcacaag cttaattaaa gcggccgcga ctctagatca taatcagcca taccacattt
     3721 gtagaggttt tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa
     3781 atgaatgcaa ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc
     3841 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     3901 tccaaactca tcaatgtatc ttaaggcgta aattgtaagc gttaatattt tgttaaaatt
     3961 cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat
     4021 cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa
     4081 gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg
     4141 cgatggccca ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa
     4201 agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc
     4261 gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag
     4321 tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg
     4381 cgcgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     4441 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     4501 ttgaaaaagg aagagtcctg aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg
     4561 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     4621 tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     4681 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact
     4741 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
     4801 gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
     4861 ctaggctttt gcaaagatcg atcaagagac aggatgagga tcgtttcgca tgattgaaca
     4921 agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg
     4981 ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg
     5041 cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aagacgaggc
     5101 agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt
     5161 cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc
     5221 atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca
     5281 tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc
     5341 acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg
     5401 gctcgcgcca gccgaactgt tcgccaggct caaggcgagc atgcccgacg gcgaggatct
     5461 cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc
     5521 tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc
     5581 tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta
     5641 cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt
     5701 ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga
     5761 gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac
     5821 gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccctagg
     5881 gggaggctaa ctgaaacacg gaaggagaca ataccggaag gaacccgcgc tatgacggca
     5941 ataaaaagac agaataaaac gcacggtgtt gggtcgtttg ttcataaacg cggggttcgg
     6001 tcccagggct ggcactctgt cgatacccca ccgagacccc attggggcca atacgcccgc
     6061 gtttcttcct tttccccacc ccacccccca agttcgggtg aaggcccagg gctcgcagcc
     6121 aacgtcgggg cggcaggccc tgccatagcc tcaggttact catatatact ttagattgat
     6181 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
     6241 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     6301 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     6361 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     6421 gtaactggct tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta
     6481 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     6541 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     6601 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     6661 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     6721 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     6781 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     6841 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     6901 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     6961 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgccat gcat
//
