LOCUS       Exported                7995 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    PJ-INPP5E
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7995)
  AUTHORS   Hammond GR, Fischer MJ, Anderson KE, Holdich J, Koteci A, Balla T, 
            Irvine RF
  TITLE     PI4P and PI(4,5)P2 Are Essential But Independent Lipid Determinants 
            of Membrane Identity.
  JOURNAL   Science. 2012 Jun 21.
  PUBMED    22722250
REFERENCE   2  (bases 1 to 7995)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7995
                     /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"
     regulatory      607..616
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             613..1287
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /note="mammalian codon-optimized"
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     primer_bind     complement(743..761)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1011..1030
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     1205..1228
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     CDS             1348..1668
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
     polyA_signal    4783..4904
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4820..4839)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4874..4893
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4911..5366)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5048..5069)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5260..5279
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5393..5497
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5499..5856
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     primer_bind     complement(5522..5542)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5707..5842
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5769..5788
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5891..6685
                     /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(5945..5964)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6555..6574
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6873..6892)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6917..6964
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      7293..7881
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7782..7801
                     /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 ccggtcgcca ccatggcctc ctccgaggac gtcatcaagg agttcatgcg cttcaaggtg
      661 cgcatggagg gctccgtgaa cggccacgag ttcgagatcg agggcgaggg cgagggccgc
      721 ccctacgagg gcacccagac cgccaagctg aaggtgacca agggcggccc cctgcccttc
      781 gcctgggaca tcctgtcccc tcagttccag tacggctcca aggcctacgt gaagcacccc
      841 gccgacatcc ccgactactt gaagctgtcc ttccccgagg gcttcaagtg ggagcgcgtg
      901 atgaacttcg aggacggcgg cgtggtgacc gtgacccagg actcctccct gcaggacggc
      961 gagttcatct acaaggtgaa gctgcgcggc accaacttcc cctccgacgg ccccgtaatg
     1021 cagaagaaga ccatgggctg ggaggcctcc accgagcgga tgtaccccga ggacggcgcc
     1081 ctgaagggcg agatcaagat gaggctgaag ctgaaggacg gcggccacta cgacgccgag
     1141 gtcaagacca cctacatggc caagaagccc gtgcagctgc ccggcgccta caagaccgac
     1201 atcaagctgg acatcacctc ccacaacgag gactacacca tcgtggaaca gtacgagcgc
     1261 gccgagggcc gccactccac cggcgcctcc ggactcagat ctcgaagcgc ggccgcggga
     1321 gcaggaggag cagctcgagc ggcgctggga gtgcaggtgg aaaccatctc cccaggagac
     1381 gggcgcacct tccccaagcg cggccagacc tgcgtggtgc actacaccgg gatgcttgaa
     1441 gatggaaaga aatttgattc ctcccgggac agaaacaagc cctttaagtt tatgctaggc
     1501 aagcaggagg tgatccgagg ctgggaagaa ggggttgccc agatgagtgt gggtcagaga
     1561 gccaaactga ctatatctcc agattatgcc tatggtgcca ctgggcaccc aggcatcatc
     1621 ccaccacatg ccactctcgt cttcgatgtg gagcttctaa aactggaaag tgctggtggt
     1681 agtgctggtg gtagtgctgg tggtagtgct ggtggtagtg ctggtggtcc gcgagctcaa
     1741 gcttcgcgat cgctagatgc gacaggtcca atagtgtacg ttcaaaatgc ggacggtatc
     1801 ttcttcaagc ttgctgaggg caaaggaact aacgatgctg ttattcactt ggccaatcaa
     1861 gatcaaggtg ttcgggtcct tggagcagag gaatttcctg tgcaaggtga agtagtaaag
     1921 attgcgtcct tgatggggtt cattaagcta aagttgaaca ggtatgccat tatcgcaaat
     1981 actgtggaag agaccggtag attcaatggc cacgttttct atagagtgtt gcaacattct
     2041 atcgtatcta ccaagtttaa ctcgagaatc gattctgaag aagccgaata tatcaagcta
     2101 ctggagttgc atttgaaaaa ttccaccttt tatttttcat acacatatga tttaacaaat
     2161 tccttacaaa gaaatgaaaa ggttggtcct gcagcctcct ggaaaaccgc tgatgaacga
     2221 ttcttttgga accattactt aactgaagat ttgagaaact ttgctcatca agatcctaga
     2281 attgactcct ttatacaacc tgttatctat gggtatgcca agacagtgga cgccgttttg
     2341 aatgccaccc ctatcgttct tggtttgatt accagacgta gtatatttag ggcgggcaca
     2401 agatacttcc gtcgtggtgt tgacaaagac ggtaacgttg gcaatttcaa tgaaactgag
     2461 caaattttac tcgctgagaa tccagagagt gaaaaaatac acgttttctc cttcttacag
     2521 acaagaggat ctgtgccaat atactgggct gaaatcaaca acttgaagta caagccaaat
     2581 cttgttcttg gagaaaactc attagatgcg acaaaaaagc attttgacca gcaaaaggag
     2641 ttatatggcg acaactactt ggttaaccta gtcaaccaaa agggccacga actacccgtg
     2701 aaagagggct atgaatcagt cgtgcacgcg ctaaacgatc cgaagattca ctacgtgtat
     2761 tttgacttcc accatgaatg tcgtaagatg caatggcata gagtgaaatt gttaattgat
     2821 cacctggaga aattaggttt atctaacgaa gatttcttcc acaaggtcat agactctaat
     2881 ggtaacaccg ttgaaattgt taatgagcaa cattccgttg taagaacaaa ctctatggat
     2941 tgtttggaca gaacaaatgt cgttcaatct gttttagccc agtgggtttt gcaaaaggag
     3001 tttgaaagtg ccgatgtcgt tgctactgga agcacttggg aagacaacgc tccattgtta
     3061 acttcttacc aaaacttatg ggctgataat gcagatgcag ttagtgtggc atattcgggc
     3121 actggagctt tgaagaccga tttcacaaga accggtaagc gtacacgtct aggtgcattc
     3181 aacgattttt tgaattcagc atcacgttat taccagaaca attggactga tggtccaaga
     3241 caggattcat acgatttatt ccttggtgga tttagaccac ataccgcttc tatcaagtcg
     3301 ccattccccg gaggtaccgc tcgaggcgca gcagcgggag caggaggagc aggtcgatcg
     3361 gatcttgcag actacaagct ccgcgcgcag ccgctcctgg tgcgggccca cagcagcctg
     3421 ggccccggcc ggccgcggag ccccctggcc tgcgacgact gttcccttcg ctcggccaaa
     3481 tcctccttca gcctcctggc gcccatccgc agcaaggacg tccgcagcag gagttacctg
     3541 gagggcagcc tcctggccag cggggccctg ttgggggcgg atgagctggc ccgctacttc
     3601 ccagaccgga acgtggcact cttcgtggcc acctggaaca tgcagggcca gaaggagctc
     3661 ccgcccagcc tggacgagtt cctgctccca gccgaggccg actatgccca ggacctgtat
     3721 gtcatcgggg tccaggaggg ctgttctgac aggcgggagt gggagactcg tctgcaggag
     3781 acgctgggcc cgcactatgt gctgctgtcc tcggcggccc acggcgtgct ctacatgtcg
     3841 ctcttcatcc gcagggacct catctggttc tgctcagagg tggagtgctc cacggtgacc
     3901 acacgcatcg tgtctcagat caagaccaag ggggccttgg gcatcagctt cacctttttt
     3961 ggcacttcct tcctcttcat cacgtcccac ttcacctcag gtgacgggaa ggtggcggag
     4021 cggctgctgg actacaccag gactgtacaa gccctggtcc tgcccagaaa tgtgcccgac
     4081 accaacccct atcgctccag cgcagcggac gtcaccaccc gcttcgatga ggtgttctgg
     4141 tttggagact tcaacttccg cctgagtggc gggcgcacag tcgtggacgc cctcctgtgc
     4201 cagggcctgg tggtggacgt gccggcgctg ctgcagcacg accagctcat ccgggagatg
     4261 cggaaagggt ccatcttcaa gggcttccag gagccggaca tccacttcct cccatcatac
     4321 aagtttgaca tcgggaagga cacgtacgac agcacctcca agcagaggac gccctcatac
     4381 acggaccgcg tcttgtacag aagccgccac aagggtgaca tctgtcctgt gagctactct
     4441 tcctgccccg ggatcaagac gtccgaccac cgccctgtgt atggcctctt ccgggtgaaa
     4501 gtgaggccgg ggcgagacaa cattccgttg gcagctggca aatttgatag agaactgtac
     4561 ttactaggaa ttaaaagacg gatttcgaag gagattcaga ggcagcaagc actacagagt
     4621 cagaactcca gcaccatcgc ctccgtttct tgaggatcca ccggatctag ataactgatc
     4681 ataatcagcc ataccacatt tgtagaggtt ttacttgctt taaaaaacct cccacacctc
     4741 cccctgaacc tgaaacataa aatgaatgca attgttgttg ttaacttgtt tattgcagct
     4801 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca
     4861 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttaacgcgt aaattgtaag
     4921 cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca
     4981 ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag
     5041 tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg
     5101 gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt
     5161 tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag
     5221 agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc
     5281 gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc
     5341 gcttaatgcg ccgctacagg gcgcgtcagg tggcactttt cggggaaatg tgcgcggaac
     5401 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     5461 ctgataaatg cttcaataat attgaaaaag gaagagtcct gaggcggaaa gaaccagctg
     5521 tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     5581 caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca
     5641 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact
     5701 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
     5761 atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag
     5821 tgaggaggct tttttggagg cctaggcttt tgcaaagatc gatcaagaga caggatgagg
     5881 atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga
     5941 gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt
     6001 ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct
     6061 gaatgaactg caagacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg
     6121 cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt
     6181 gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc
     6241 tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc
     6301 gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga
     6361 tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgag
     6421 catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat
     6481 ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg
     6541 ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc
     6601 tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta
     6661 tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg
     6721 acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc
     6781 ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg
     6841 gagttcttcg cccaccctag ggggaggcta actgaaacac ggaaggagac aataccggaa
     6901 ggaacccgcg ctatgacggc aataaaaaga cagaataaaa cgcacggtgt tgggtcgttt
     6961 gttcataaac gcggggttcg gtcccagggc tggcactctg tcgatacccc accgagaccc
     7021 cattggggcc aatacgcccg cgtttcttcc ttttccccac cccacccccc aagttcgggt
     7081 gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc ctcaggttac
     7141 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     7201 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     7261 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     7321 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     7381 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc
     7441 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     7501 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     7561 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     7621 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     7681 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     7741 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     7801 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     7861 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     7921 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
     7981 attaccgcca tgcat
//
