LOCUS       sequence-419031-        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: Pseudojanin (PJ)
FEATURES             Location/Qualifiers
     source          1..7995
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     40..59
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(77..532)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(214..235)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     426..445
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        559..663
                     /gene="bla"
                     /label=AmpR promoter
     promoter        665..1022
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        665..856
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(688..708)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      873..1008
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             1057..1851
                     /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             1057..1851
                     /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(1111..1130)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1721..1740
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(2039..2058)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    2083..2130
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      2459..3047
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2948..2967
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        3222..3525
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3526..3729
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3680..3700
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      3768..3777
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3774..4448
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     CDS             3774..4448
                     /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(3904..3922)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     4172..4191
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     4366..4389
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     CDS             4509..4829
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
     polyA_signal    join(7944..7995,1..70)
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(join(7981..7995,1..5))
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
ORIGIN
        1 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
       61 atgtatctta acgcgtaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt
      121 tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca
      181 aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta
      241 aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta
      301 cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg
      361 aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga
      421 aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg
      481 ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcaggtggc
      541 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
      601 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
      661 agtcctgagg cggaaagaac cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc
      721 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt
      781 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
      841 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg
      901 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
      961 cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca
     1021 aagatcgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac
     1081 gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca
     1141 atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt
     1201 gtcaagaccg acctgtccgg tgccctgaat gaactgcaag acgaggcagc gcggctatcg
     1261 tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga
     1321 agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct
     1381 cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg
     1441 gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg
     1501 gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc
     1561 gaactgttcg ccaggctcaa ggcgagcatg cccgacggcg aggatctcgt cgtgacccat
     1621 ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac
     1681 tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt
     1741 gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct
     1801 cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc
     1861 tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca
     1921 ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga
     1981 tcctccagcg cggggatctc atgctggagt tcttcgccca ccctaggggg aggctaactg
     2041 aaacacggaa ggagacaata ccggaaggaa cccgcgctat gacggcaata aaaagacaga
     2101 ataaaacgca cggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc
     2161 actctgtcga taccccaccg agaccccatt ggggccaata cgcccgcgtt tcttcctttt
     2221 ccccacccca ccccccaagt tcgggtgaag gcccagggct cgcagccaac gtcggggcgg
     2281 caggccctgc catagcctca ggttactcat atatacttta gattgattta aaacttcatt
     2341 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     2401 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     2461 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     2521 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     2581 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     2641 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     2701 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     2761 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     2821 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     2881 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     2941 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     3001 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     3061 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
     3121 tatcccctga ttctgtggat aaccgtatta ccgccatgca ttagttatta atagtaatca
     3181 attacggggt cattagttca tagcccatat atggagttcc gcgttacata acttacggta
     3241 aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat
     3301 gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg
     3361 taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc ccctattgac
     3421 gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt atgggacttt
     3481 cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtgat gcggttttgg
     3541 cagtacatca atgggcgtgg atagcggttt gactcacggg gatttccaag tctccacccc
     3601 attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc aaaatgtcgt
     3661 aacaactccg ccccattgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata
     3721 agcagagctg gtttagtgaa ccgtcagatc cgctagcgct accggtcgcc accatggcct
     3781 cctccgagga cgtcatcaag gagttcatgc gcttcaaggt gcgcatggag ggctccgtga
     3841 acggccacga gttcgagatc gagggcgagg gcgagggccg cccctacgag ggcacccaga
     3901 ccgccaagct gaaggtgacc aagggcggcc ccctgccctt cgcctgggac atcctgtccc
     3961 ctcagttcca gtacggctcc aaggcctacg tgaagcaccc cgccgacatc cccgactact
     4021 tgaagctgtc cttccccgag ggcttcaagt gggagcgcgt gatgaacttc gaggacggcg
     4081 gcgtggtgac cgtgacccag gactcctccc tgcaggacgg cgagttcatc tacaaggtga
     4141 agctgcgcgg caccaacttc ccctccgacg gccccgtaat gcagaagaag accatgggct
     4201 gggaggcctc caccgagcgg atgtaccccg aggacggcgc cctgaagggc gagatcaaga
     4261 tgaggctgaa gctgaaggac ggcggccact acgacgccga ggtcaagacc acctacatgg
     4321 ccaagaagcc cgtgcagctg cccggcgcct acaagaccga catcaagctg gacatcacct
     4381 cccacaacga ggactacacc atcgtggaac agtacgagcg cgccgagggc cgccactcca
     4441 ccggcgcctc cggactcaga tctcgaagcg cggccgcggg agcaggagga gcagctcgag
     4501 cggcgctggg agtgcaggtg gaaaccatct ccccaggaga cgggcgcacc ttccccaagc
     4561 gcggccagac ctgcgtggtg cactacaccg ggatgcttga agatggaaag aaatttgatt
     4621 cctcccggga cagaaacaag ccctttaagt ttatgctagg caagcaggag gtgatccgag
     4681 gctgggaaga aggggttgcc cagatgagtg tgggtcagag agccaaactg actatatctc
     4741 cagattatgc ctatggtgcc actgggcacc caggcatcat cccaccacat gccactctcg
     4801 tcttcgatgt ggagcttcta aaactggaaa gtgctggtgg tagtgctggt ggtagtgctg
     4861 gtggtagtgc tggtggtagt gctggtggtc cgcgagctca agcttcgcga tcgctagatg
     4921 cgacaggtcc aatagtgtac gttcaaaatg cggacggtat cttcttcaag cttgctgagg
     4981 gcaaaggaac taacgatgct gttattcact tggccaatca agatcaaggt gttcgggtcc
     5041 ttggagcaga ggaatttcct gtgcaaggtg aagtagtaaa gattgcgtcc ttgatggggt
     5101 tcattaagct aaagttgaac aggtatgcca ttatcgcaaa tactgtggaa gagaccggta
     5161 gattcaatgg ccacgttttc tatagagtgt tgcaacattc tatcgtatct accaagttta
     5221 actcgagaat cgattctgaa gaagccgaat atatcaagct actggagttg catttgaaaa
     5281 attccacctt ttatttttca tacacatatg atttaacaaa ttccttacaa agaaatgaaa
     5341 aggttggtcc tgcagcctcc tggaaaaccg ctgatgaacg attcttttgg aaccattact
     5401 taactgaaga tttgagaaac tttgctcatc aagatcctag aattgactcc tttatacaac
     5461 ctgttatcta tgggtatgcc aagacagtgg acgccgtttt gaatgccacc cccatcgttc
     5521 ttggtttgat taccagacgt agtatattta gggcgggcac aagatacttc cgtcgtggtg
     5581 ttgacaaaga cggtaacgtt ggcaatttca atgaaactga gcaaatttta ctcgctgaga
     5641 atccagagag tgaaaaaata cacgttttct ccttcttaca gacaagagga tctgtgccaa
     5701 tatactgggc tgaaatcaac aacttgaagt acaagccaaa tcttgttctt ggagaaaact
     5761 cattagatgc gacaaaaaag cattttgacc agcaaaagga gttatatggc gacaactact
     5821 tggttaacct agtcaaccaa aagggccacg aactacccgt gaaagagggc tatgaatcag
     5881 ttgtgcacgc gctaaacgat ccgaagattc actacgtgta ttttgacttc caccatgaat
     5941 gtcgtaagat gcaatggcat agagtgaaac tgttaattga tcacttggag aaattaggtt
     6001 tatctaacga agatttcttc cacaaggtca tagactctaa tggtaacacc gttgaaattg
     6061 ttaatgagca acattccgtt gtaagaacaa actgtatgga ttgtttggac agaacaaatg
     6121 tcgttcaatc tgttctagcc cagtgggttt tgcaaaagga gtttgaaagt gccgatgtcg
     6181 ttgctactgg aagcacttgg gaagacaacg ctccattgtt aacttcttac caaaacttat
     6241 gggctgataa cgcagatgca gttagtgtgg catattcggg cactggagct ttgaagaccg
     6301 atttcacaag aaccggtaag cgtacacgtc taggtgcatt caacgatttt ttgaattcag
     6361 catcacgtta ttaccagaac aattggactg atggtccaag acaggattca tacgatttat
     6421 tccttggtgg atttagacca cataccgctt ctatcaagtc gccattcccc ggaggtaccg
     6481 ctcgaggcgc agcagcggga gcaggaggag caggtcgatc ggatcttgca gactacaagc
     6541 tccgcgcgca gccgctcctg gtgcgggccc acagcagcct gggccccggc cggccgcgga
     6601 gccccctggc ctgcgacgac tgttcccttc gctcggccaa atcctccttc agcctcctgg
     6661 cgcccatccg cagcaaggac gtccgcagca ggagttacct ggagggcagc ctcctggcca
     6721 gcggggccct gttgggggcg gatgagctgg cccgctactt cccagaccgg aacgtggcac
     6781 tcttcgtggc cacctggaac atgcagggcc agaaggagct cccgcccagc ctggacgagt
     6841 tcctgctccc agccgaggcc gactatgccc aggacctgta tgtcatcggg gtccaggagg
     6901 gctgttctga caggcgggag tgggagactc gtctgcagga gacgctgggc ccgcactatg
     6961 tgctgctgtc ctcggcggcc cacggcgtgc tctacatgtc gctcttcatc cgcagggacc
     7021 tcatctggtt ctgctcagag gtggagtgct ccacggtgac cacacgcatc gtgtctcaga
     7081 tcaagaccaa gggggccttg ggcatcagct tcaccttttt tggcacttcc ttcctcttca
     7141 tcacgtccca cttcacctca ggtgacggga aggtggcgga gcggctgctg gactacacca
     7201 ggactgtaca agccctggtc ctgcccagaa atgtgcccga caccaacccc tatcgctcca
     7261 gcgcagcgga cgtcaccacc cgcttcgatg aggtgttctg gtttggagac ttcaacttcc
     7321 gcctgagtgg cgggcgcaca gtcgtggacg ccctcctgtg ccagggcctg gtggtggacg
     7381 tgccggcgct gctgcagcac gaccagctca tccgggagat gcggaaaggg tccatcttca
     7441 agggcttcca ggagccggac atccacttcc tcccatcata caagtttgac atcgggaagg
     7501 acacgtacga cagcacctcc aagcagagga cgccctcata cacggaccgc gtcttgtaca
     7561 gaagccgcca caagggtgac atctgtcctg tgagctactc ttcctgcccc gggatcaaga
     7621 cgtccgacca ccgccctgtg tatggcctct tccgggtgaa agtgaggccg gggcgagaca
     7681 acattccgtt ggcagctggc aaatttgata gagaactgta cttactagga attaaaagac
     7741 ggatttcgaa ggagattcag aggcagcaag cactacagag tcagaactcc agcaccatcg
     7801 cctccgtttc ttgaggatcc accggatcta gataactgat cataatcagc cataccacat
     7861 ttgtagaggt tttacttgct ttaaaaaacc tcccacacct ccccctgaac ctgaaacata
     7921 aaatgaatgc aattgttgtt gttaacttgt ttattgcagc ttataatggt tacaaataaa
     7981 gcaatagcat cacaa
//
