LOCUS       Exported                8904 bp ds-DNA     circular SYN 25-DEC-2023
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    pIRES-split-Cdc42-T17N
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8904)
  AUTHORS   gene
  TITLE     Direct Submission
  JOURNAL   Exported Tuesday, Jan 2, 2024 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8904
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        138..517
                     /note="CMV enhancer"
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        518..729
                     /note="CMV promoter"
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     intron          890..1022
                     /note="chimeric intron"
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     promoter        1067..1085
                     /note="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1091..1807
                     /codon_start=1
                     /product="enhanced monomeric variant of CFP (Rizzo et al.,
                     2004)"
                     /label=enhanced monomeric variant of CFP (Rizzo et al
                     /note="mCerulean"
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNAISDNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             1832..2101
                     /codon_start=1
                     /product="FKBP-rapamycin binding domain of human FRAP"
                     /label=FKBP-rapamycin binding domain of human FRAP
                     /note="FRB"
                     /translation="EMWHEGLEEASRLYFGERNVKGMFEVLEPLHAMMERGPQTLKETS
                     FNQAYGRDLMEAQEWCRKYMKSGNVKDLTQAWDLYYHVFRRISKQ"
     misc_feature    2248..2821
                     /label=IRES
                     /note="IRES"
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     CDS             3458..3778
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=human FK506-binding protein FKBP12
                     /note="FKBP"
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
     CDS             3803..4516
                     /codon_start=1
                     /product="Venus YFP with monomerizing A206K mutation (Nagai
                     et al., 2002)"
                     /label=Venus YFP with monomerizing A206K mutation (Nag
                     /note="mVenus"
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     LICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKA
                     NFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     promoter        complement(4573..4590)
                     /note="T3 promoter"
                     /note="promoter for bacteriophage T3 RNA polymerase"
     polyA_signal    4608..4729
                     /label=SV40 poly(A) signal
                     /note="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     rep_origin      4915..5370
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        5487..5844
                     /note="SV40 promoter"
                     /note="SV40 enhancer and early promoter"
     rep_origin      5695..5830
                     /label=SV40 ori
                     /note="SV40 ori"
                     /note="SV40 origin of replication"
     CDS             5895..6689
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=aph(3')-II (or nptII)
                     /note="NeoR/KanR"
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    6753..6801
                     /label=synthetic polyadenylation signal
                     /note="synthetic polyadenylation signal"
     promoter        7107..7211
                     /gene="bla"
                     /label=bla promoter
                     /note="AmpR promoter"
     CDS             7212..8072
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=bla
                     /note="AmpR"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      8243..8831
                     /direction=RIGHT
                     /label=ori
                     /note="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 tcaatattgg ccattagcca tattattcat tggttatata gcataaatca atattggcta
       61 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
      121 aatatgaccg ccatgttggc attgattatt gactagttat taatagtaat caattacggg
      181 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      241 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      301 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      361 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
      421 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
      481 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
      541 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      601 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactg
      661 cgatcgcccg ccccgttgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata
      721 agcagagctc gtttagtgaa ccgtcagatc actagaagct ttattgcggt agtttatcac
      781 agttaaattg ctaacgcagt cagtgcttct gacacaacag tctcgaactt aagctgcagt
      841 gactctctta aggtagcctt gcagaagttg gtcgtgaggc actgggcagg taagtatcaa
      901 ggttacaaga caggtttaag gagaccaata gaaactgggc ttgtcgagac agagaagact
      961 cttgcgtttc tgataggcac ctattggtct tactgacatc cactttgcct ttctctccac
     1021 aggtgtccac tcccagttca attacagctc ttaaggctag agtacttaat acgactcact
     1081 ataggctagc atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt
     1141 cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga
     1201 tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc
     1261 ctggcccacc ctcgtgacca ccctgacctg gggcgtgcag tgcttcgccc gctaccccga
     1321 ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg
     1381 caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg
     1441 cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat
     1501 cctggggcac aagctggagt acaacgccat cagcgacaac gtctatatca ccgccgacaa
     1561 gcagaagaac ggcatcaagg ccaacttcaa gatccgccac aacatcgagg acggcagcgt
     1621 gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc
     1681 cgacaaccac tacctgagca cccagtccaa gctgagcaaa gaccccaacg agaagcgcga
     1741 tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct
     1801 gtacaagggc ggtggctcat ctggcggagg tgagatgtgg catgaaggcc tggaagaggc
     1861 atctcgtttg tactttgggg aaaggaacgt gaaaggcatg tttgaggtgc tggagccctt
     1921 gcatgctatg atggaacggg gcccccagac tctgaaggaa acatccttta atcaggccta
     1981 tggtcgagat ttaatggagg cccaagagtg gtgcaggaag tacatgaaat cagggaatgt
     2041 caaggacctc acccaagcct gggacctcta ttatcatgtg ttccgacgaa tctcaaagca
     2101 gcagatctcg tacgcgtccc ggggcggtgg ctcatctggc ggaggtgagc tccagacaat
     2161 taagtgtgtt gttgtgggcg atggtgctta agaattcacg cgtcgagcat gcatctaggg
     2221 cggccaattc cgcccctctc cccccccccc ctctccctcc ccccccccta acgttactgg
     2281 ccgaagccgc ttggaataag gccggtgtgc gtttgtctat atgttatttt ccaccatatt
     2341 gccgtctttt ggcaatgtga gggcccggaa acctggccct gtcttcttga cgagcattcc
     2401 taggggtctt tcccctctcg ccaaaggaat gcaaggtctg ttgaatgtcg tgaaggaagc
     2461 agttcctctg gaagcttctt gaagacaaac aacgtctgta gcgacccttt gcaggcagcg
     2521 gaacccccca cctggcgaca ggtgcctctg cggccaaaag ccacgtgtat aagatacacc
     2581 tgcaaaggcg gcacaacccc agtgccacgt tgtgagttgg atagttgtgg aaagagtcaa
     2641 atggctctcc tcaagcgtat tcaacaaggg gctgaaggat gcccagaagg taccccattg
     2701 tatgggatct gatctggggc ctcggtgcac atgctttaca tgtgtttagt cgaggttaaa
     2761 aaaacgtcta ggccccccga accacgggga cgtggttttc ctttgaaaaa cacgatgata
     2821 agcttgccac aacccgggat cctctagaat gggggttggt aaaaactgtc tcctgatatc
     2881 ctacacaaca aacaaatttc catcggagta tgtaccgact gtttttgaca actatgcagt
     2941 cacagttatg attggtggag aaccatatac tcttggactt tttgatactg cagggcaaga
     3001 ggattatgac agattacgac cgctgagtta tccacaaaca gatgtatttc tagtctgttt
     3061 ttcagtggtc tctccatctt catttgaaaa cgtgaaagaa aagtgggtgc ctgagataac
     3121 tcaccactgt ccaaagactc ctttcttgct tgttgggact caaattgatc tcagagatga
     3181 cccctctact attgagaaac ttgccaagaa caaacagaag cctatcactc cagagactgc
     3241 tgaaaagctg gcccgtgacc tgaaggctgt caagtatgtg gagtgttctg cacttacaca
     3301 gagaggtctg aagaatgtgt ttgatgaggc tatcctagct gccctcgagc ctccggaaac
     3361 tcaacccggg gatcccaatt gggagctcgt gtacacggcg cgcctgcagg gaggtggctc
     3421 atctggcgga ggtcagatct cgtacgcgtc ccggggcgga gtgcaggtgg aaaccatctc
     3481 cccaggagac gggcgcacct tccccaagcg cggccagacc tgcgtggtgc actacaccgg
     3541 gatgcttgaa gatggaaaga aatttgattc ctcccgggac agaaacaagc cctttaagtt
     3601 tatgctaggc aagcaggagg tgatccgagg ctgggaagaa ggggttgccc agatgagtgt
     3661 gggtcagaga gccaaactga ctatatctcc agattatgcc tatggtgcca ctgggcaccc
     3721 aggcatcatc ccaccacatg ccactctcgt cttcgatgtg gagcttctaa aactggaagg
     3781 cggtggctca tctggcggag gtgtgagcaa gggcgaggag ctgttcaccg gggtggtgcc
     3841 catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg
     3901 cgagggcgat gccacctacg gcaagctgac cctgaagctc atctgcacca ccggcaagct
     3961 gcccgtgccc tggcccaccc tcgtgaccac cctcggctac ggcctgcagt gcttcgcccg
     4021 ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt
     4081 ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa
     4141 gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga
     4201 cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg tctatatcac
     4261 cgccgacaag cagaagaacg gcatcaaggc caacttcaag atccgccaca acatcgagga
     4321 cggcggcgtg cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt
     4381 gctgctgccc gacaaccact acctgagcta ccagtccaag ctgagcaaag accccaacga
     4441 gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat
     4501 ggacgagctg tacaagaaaa aaaagaagaa aaagagcaag accaaatgcg tgattatgta
     4561 agcggccgct tccctttagt gagggttaat gcttcgagca gacatgataa gatacattga
     4621 tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg
     4681 tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagtta acaacaacaa
     4741 ttgcattcat tttatgtttc aggttcaggg ggagatgtgg gaggtttttt aaagcaagta
     4801 aaacctctac aaatgtggta aaatccgata aggatcgatc cgggctggcg taatagcgaa
     4861 gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga atggacgcgc
     4921 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     4981 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     5041 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
     5101 tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc
     5161 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
     5221 tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga
     5281 ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
     5341 attttaacaa aatattaacg cttacaattt cctgatgcgg tattttctcc ttacgcatct
     5401 gtgcggtatt tcacaccgca tacgcggatc tgcgcagcac catggcctga aataacctct
     5461 gaaagaggaa cttggttagg taccttctga ggcggaaaga accagctgtg gaatgtgtgt
     5521 cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     5581 ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     5641 caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg
     5701 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
     5761 tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt
     5821 tttggaggcc taggcttttg caaaaagctt gattcttctg acacaacagt ctcgaactta
     5881 aggctagagc caccatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg
     5941 tggagaggct attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg
     6001 tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg
     6061 ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc
     6121 cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg
     6181 aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca
     6241 tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc
     6301 aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg
     6361 atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg
     6421 cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata
     6481 tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg
     6541 accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat
     6601 gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct
     6661 tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca
     6721 agcgacgccc aacctgccat cacgatggcc gcaataaaat atctttattt tcattacatc
     6781 tgtgtgttgg ttttttgtgt gaatcgatag cgataaggat ccgcgtatgg tgcactctca
     6841 gtacaatctg ctctgatgcc gcatagttaa gccagccccg acacccgcca acacccgctg
     6901 acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct
     6961 ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg
     7021 gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt
     7081 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     7141 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     7201 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     7261 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     7321 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     7381 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
     7441 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     7501 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
     7561 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     7621 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     7681 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     7741 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
     7801 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
     7861 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     7921 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     7981 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     8041 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
     8101 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     8161 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     8221 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     8281 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     8341 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     8401 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     8461 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     8521 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     8581 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     8641 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     8701 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     8761 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     8821 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     8881 ttgctcacat ggctcgacag atct
//
