LOCUS       sequence-352654-        7995 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 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 Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Science. 
            2012 Jun 21."
COMMENT     Sequence Label: PJ-Sac
FEATURES             Location/Qualifiers
     source          1..7995
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        128..319
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     276..296
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(321..425)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      452..907
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(539..558)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     749..770
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    914..1035
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(925..944)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     979..998
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(4150..4470)
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
     CDS             complement(4531..5205)
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     CDS             complement(4531..5205)
                     /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(4590..4613)
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     primer_bind     complement(4788..4807)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     5057..5075
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     regulatory      5202..5211
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(5250..5453)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(5279..5299)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        5454..5757
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(5932..6520)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6012..6031)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    6849..6896
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     6921..6940
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(7128..7922)
                     /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             complement(7128..7922)
                     /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(7239..7258)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     7849..7868
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(join(7957..7995,1..319))
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      join(7971..7995,1..111)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 cttctggaat agctcagagg ccgaggcggc ctcggcctct gcataaataa aaaaaattag
       61 tcagccatgg ggcggagaat gggcggaact gggcggagtt aggggcggga tgggcggagt
      121 taggggcggg actatggttg ctgactaatt gagatgcatg ctttgcatac ttctgcctgc
      181 tggggagcct ggggactttc cacacctggt tgctgactaa ttgagatgca tgctttgcat
      241 acttctgcct gctggggagc ctggggactt tccacaccct aactgacaca cattccacag
      301 ctggttcttt ccgcctcagg actcttcctt tttcaatatt attgaagcat ttatcagggt
      361 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
      421 ccgcgcacat ttccccgaaa agtgccacct gacgcgccct gtagcggcgc attaagcgcg
      481 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
      541 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
      601 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
      661 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
      721 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
      781 aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg
      841 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgctt
      901 acaatttacg cgttaagata cattgatgag tttggacaaa ccacaactag aatgcagtga
      961 aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac cattataagc
     1021 tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt tcagggggag
     1081 gtgtgggagg ttttttaaag caagtaaaac ctctacaaat gtggtatggc tgattatgat
     1141 cagttatcta gatccggtgg atcctcaaga aacggaggcg atggtgctgg agttctgact
     1201 ctgtagtgct tgctgcctct gaatctcctt cgaaatccgt cttttaattc ctagtaagta
     1261 cagttctcta tcaaatttgc cagctgccaa cggaatgttg tctcgccccg gcctcacttt
     1321 cacccggaag aggccataca cagggcggtg gtcggacgtc ttgatcccgg ggcaggaaga
     1381 gtagctcaca ggacagatgt cacccttgtg gcggcttctg tacaagacgc gggccgtgta
     1441 tgagggcgtc ctctgcttgg aggtgctgtc gtacgtgtcc ttcccgatgt caaacttgta
     1501 tgatgggagg aagtggatgt ccggctcctg gaagcccttg aagatggacc ctttccgcat
     1561 ctcccggatg agctggtcgt gctgcagcag cgccggcacg tccaccacca ggccctggca
     1621 caggagggcg tccacgactg tgcgcccgcc actcaggcgg aagttgaagt ctccaaacca
     1681 gaacacctca tcgaagcggg tggtgacgtc cgctgcgctg gagcgatagg ggttggtgtc
     1741 gggcacattt ctgggcagga ccagggcttg tacagtcctg gtgtagtcca gcagccgctc
     1801 cgccaccttc ccgtcacctg aggtgaagtg ggacgtgatg aagaggaagg aagtgccaaa
     1861 aaaggtgaag ctgatgccca aggccccctt ggtcttgatc tgagacacga tgcgtgtggt
     1921 caccgtggag cactccacct ctgagcagaa ccagatgagg tccctgcgga tgaagagcga
     1981 catgtagagc acgccgtggg ccgccgagga cagcagcaca tagtgcgggc ccagcgtctc
     2041 ctgcagacga gtctcccact cccgcctgtc agaacagccc tcctggaccc cgatgacata
     2101 caggtcctgg gcatagtcgg cctcggctgg gagcaggaac tcgtccaggc tgggcgggag
     2161 ctccttctgg ccctgcatgt tccaggtggc cacgaagagt gccacgttcc ggtctgggaa
     2221 gtagcgggcc agctcatccg cccccaacag ggccccgctg gccaggaggc tgccctccag
     2281 gtaactcctg ctgcggacgt ccttgctgcg gatgggcgcc aggaggctga aggaggattt
     2341 ggccgagcga agggaacagt cgtcgcaggc cagggggctc cgcggccggc cggggcccag
     2401 gctgctgtgg gcccgcacca ggagcggctg cgcgcggagc ttgtagtctg caagatccga
     2461 tcgacctgct cctcctgctc ccgctgctgc gcctcgagcg gtacctccgg ggaatggcga
     2521 cttgatagaa gcggtatgtg gtctaaatcc accaaggaat aaatcgtatg aatcctgtct
     2581 tggaccatca gtccaattgt tctggtaata acgtgatgct gaattcaaaa aatcgttgaa
     2641 tgcacctaga cgtgtacgct taccggttct tgtgaaatcg gtcttcaaag ctccagtgcc
     2701 cgaatatgcc acactaactg catctgcgtt atcagcccat aagttttggt aagaagttaa
     2761 caatggagcg ttgtcttccc aagtgcttcc agtagcaacg acatcggcac tttcaaactc
     2821 cttttgcaaa acccactggg ctagaacaga ttgaacgaca tttgttctgt ccaaacaatc
     2881 catacagttt gttcttacaa cggaatgttg ctcattaaca atttcaacgg tgttaccatt
     2941 agagtctatg accttgtgga agaaatcttc gttagataaa cctaatttct ccaagtgatc
     3001 aattaacagt ttcactctat gccattgcat cttacgacat tcatggtgga agtcaaaata
     3061 cacgtagtga atcttcggat cgtttagcgc gtgcacaact gattcatagc cctctttcac
     3121 gggtagttcg tggccctttt ggttgactag gttaaccaag tagttgtcgc catataactc
     3181 cttttgctgg tcaaaatgct tttttgtcgc atctaatgag ttttctccaa gaacaagatt
     3241 tggcttgtac ttcaagttgt tgatttcagc ccagtatatt ggcacagatc ctcttgtctg
     3301 taagaaggag aaaacgtgta ttttttcact ctctggattc tcagcgagta aaatttgctc
     3361 agtttcattg aaattgccaa cgttaccgtc tttgtcaaca ccacgacgga agtatcttgt
     3421 gcccgcccta aatatactac gtctggtaat caaaccaaga acgatggggg tggcattcaa
     3481 aacggcgtcc actgtcttgg catacccata gataacaggt tgtataaagg agtcaattct
     3541 aggatcttga tgagcaaagt ttctcaaatc ttcagttaag taatggttcc aaaagaatcg
     3601 ttcatcagcg gttttccagg aggctgcagg accaaccttt tcatttcttt gtaaggaatt
     3661 tgttaaatca tatgtgtatg aaaaataaaa ggtggaattt ttcaaatgca actccagtag
     3721 cttgatatat tcggcttctt cagaatcgat tctcgagtta aacttggtag atacgataga
     3781 atgttgcaac actctataga aaacgtggcc attgaatcta ccggtctctt ccacagtatt
     3841 tgcgataatg gcatacctgt tcaactttag cttaatgaac cccatcaagg acgcaatctt
     3901 tactacttca ccttgcacag gaaattcctc tgctccaagg acccgaacac cttgatcttg
     3961 attggccaag tgaataacag catcgttagt tcctttgccc tcagcaagct tgaagaagat
     4021 accgtccgca ttttgaacgt atactattgg acctgtcgca tctagcgatc gcgaagcttg
     4081 agctcgcgga ccaccagcac taccaccagc actaccacca gcactaccac cagcactacc
     4141 accagcactt tccagtttta gaagctccac atcgaagacg agagtggcat gtggtgggat
     4201 gatgcctggg tgcccagtgg caccataggc ataatctgga gatatagtca gtttggctct
     4261 ctgacccaca ctcatctggg caaccccttc ttcccagcct cggatcacct cctgcttgcc
     4321 tagcataaac ttaaagggct tgtttctgtc ccgggaggaa tcaaatttct ttccatcttc
     4381 aagcatcccg gtgtagtgca ccacgcaggt ctggccgcgc ttggggaagg tgcgcccgtc
     4441 tcctggggag atggtttcca cctgcactcc cagcgccgct cgagctgctc ctcctgctcc
     4501 cgcggccgcg cttcgagatc tgagtccgga ggcgccggtg gagtggcggc cctcggcgcg
     4561 ctcgtactgt tccacgatgg tgtagtcctc gttgtgggag gtgatgtcca gcttgatgtc
     4621 ggtcttgtag gcgccgggca gctgcacggg cttcttggcc atgtaggtgg tcttgacctc
     4681 ggcgtcgtag tggccgccgt ccttcagctt cagcctcatc ttgatctcgc ccttcagggc
     4741 gccgtcctcg gggtacatcc gctcggtgga ggcctcccag cccatggtct tcttctgcat
     4801 tacggggccg tcggagggga agttggtgcc gcgcagcttc accttgtaga tgaactcgcc
     4861 gtcctgcagg gaggagtcct gggtcacggt caccacgccg ccgtcctcga agttcatcac
     4921 gcgctcccac ttgaagccct cggggaagga cagcttcaag tagtcgggga tgtcggcggg
     4981 gtgcttcacg taggccttgg agccgtactg gaactgaggg gacaggatgt cccaggcgaa
     5041 gggcaggggg ccgcccttgg tcaccttcag cttggcggtc tgggtgccct cgtaggggcg
     5101 gccctcgccc tcgccctcga tctcgaactc gtggccgttc acggagccct ccatgcgcac
     5161 cttgaagcgc atgaactcct tgatgacgtc ctcggaggag gccatggtgg cgaccggtag
     5221 cgctagcgga tctgacggtt cactaaacca gctctgctta tatagacctc ccaccgtaca
     5281 cgcctaccgc ccatttgcgt caatggggcg gagttgttac gacattttgg aaagtcccgt
     5341 tgattttggt gccaaaacaa actcccattg acgtcaatgg ggtggagact tggaaatccc
     5401 cgtgagtcaa accgctatcc acgcccattg atgtactgcc aaaaccgcat caccatggta
     5461 atagcgatga ctaatacgta gatgtactgc caagtaggaa agtcccataa ggtcatgtac
     5521 tgggcataat gccaggcggg ccatttaccg tcattgacgt caataggggg cgtacttggc
     5581 atatgataca cttgatgtac tgccaagtgg gcagtttacc gtaaatactc cacccattga
     5641 cgtcaatgga aagtccctat tggcgttact atgggaacat acgtcattat tgacgtcaat
     5701 gggcgggggt cgttgggcgg tcagccaggc gggccattta ccgtaagtta tgtaacgcgg
     5761 aactccatat atgggctatg aactaatgac cccgtaattg attactatta ataactaatg
     5821 catggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     5881 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     5941 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     6001 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     6061 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     6121 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     6181 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     6241 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     6301 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     6361 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     6421 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     6481 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     6541 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     6601 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     6661 aagtatatat gagtaacctg aggctatggc agggcctgcc gccccgacgt tggctgcgag
     6721 ccctgggcct tcacccgaac ttggggggtg gggtggggaa aaggaagaaa cgcgggcgta
     6781 ttggccccaa tggggtctcg gtggggtatc gacagagtgc cagccctggg accgaacccc
     6841 gcgtttatga acaaacgacc caacaccgtg cgttttattc tgtcttttta ttgccgtcat
     6901 agcgcgggtt ccttccggta ttgtctcctt ccgtgtttca gttagcctcc ccctagggtg
     6961 ggcgaagaac tccagcatga gatccccgcg ctggaggatc atccagccgg cgtcccggaa
     7021 aacgattccg aagcccaacc tttcatagaa ggcggcggtg gaatcgaaat ctcgtgatgg
     7081 caggttgggc gtcgcttggt cggtcatttc gaaccccaga gtcccgctca gaagaactcg
     7141 tcaagaaggc gatagaaggc gatgcgctgc gaatcgggag cggcgatacc gtaaagcacg
     7201 aggaagcggt cagcccattc gccgccaagc tcttcagcaa tatcacgggt agccaacgct
     7261 atgtcctgat agcggtccgc cacacccagc cggccacagt cgatgaatcc agaaaagcgg
     7321 ccattttcca ccatgatatt cggcaagcag gcatcgccat gggtcacgac gagatcctcg
     7381 ccgtcgggca tgctcgcctt gagcctggcg aacagttcgg ctggcgcgag cccctgatgc
     7441 tcttcgtcca gatcatcctg atcgacaaga ccggcttcca tccgagtacg tgctcgctcg
     7501 atgcgatgtt tcgcttggtg gtcgaatggg caggtagccg gatcaagcgt atgcagccgc
     7561 cgcattgcat cagccatgat ggatactttc tcggcaggag caaggtgaga tgacaggaga
     7621 tcctgccccg gcacttcgcc caatagcagc cagtcccttc ccgcttcagt gacaacgtcg
     7681 agcacagctg cgcaaggaac gcccgtcgtg gccagccacg atagccgcgc tgcctcgtct
     7741 tgcagttcat tcagggcacc ggacaggtcg gtcttgacaa aaagaaccgg gcgcccctgc
     7801 gctgacagcc ggaacacggc ggcatcagag cagccgattg tctgttgtgc ccagtcatag
     7861 ccgaatagcc tctccaccca agcggccgga gaacctgcgt gcaatccatc ttgttcaatc
     7921 atgcgaaacg atcctcatcc tgtctcttga tcgatctttg caaaagccta ggcctccaaa
     7981 aaagcctcct cacta
//
