LOCUS       Exported                8892 bp ds-DNA     circular SYN 25-DEC-2023
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    pIRES-split-RhoA (T19N)
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8892)
  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..8892
                     /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    2254..2827
                     /label=IRES
                     /note="IRES"
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     CDS             3446..3766
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=human FK506-binding protein FKBP12
                     /note="FKBP"
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
     CDS             3791..4504
                     /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(4561..4578)
                     /note="T3 promoter"
                     /note="promoter for bacteriophage T3 RNA polymerase"
     polyA_signal    4596..4717
                     /label=SV40 poly(A) signal
                     /note="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     rep_origin      4903..5358
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        5475..5832
                     /note="SV40 promoter"
                     /note="SV40 enhancer and early promoter"
     rep_origin      5683..5818
                     /label=SV40 ori
                     /note="SV40 ori"
                     /note="SV40 origin of replication"
     CDS             5883..6677
                     /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    6741..6789
                     /label=synthetic polyadenylation signal
                     /note="synthetic polyadenylation signal"
     promoter        7095..7199
                     /gene="bla"
                     /label=bla promoter
                     /note="AmpR promoter"
     CDS             7200..8060
                     /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      8231..8819
                     /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 tcgctgccat
     2161 ccggaagaaa ctggtgattg ttggtgatgg agcctaagaa ttcacgcgtc gagcatgcat
     2221 ctagggcggc caattccgcc cctctccccc ccccccctct ccctcccccc cccctaacgt
     2281 tactggccga agccgcttgg aataaggccg gtgtgcgttt gtctatatgt tattttccac
     2341 catattgccg tcttttggca atgtgagggc ccggaaacct ggccctgtct tcttgacgag
     2401 cattcctagg ggtctttccc ctctcgccaa aggaatgcaa ggtctgttga atgtcgtgaa
     2461 ggaagcagtt cctctggaag cttcttgaag acaaacaacg tctgtagcga ccctttgcag
     2521 gcagcggaac cccccacctg gcgacaggtg cctctgcggc caaaagccac gtgtataaga
     2581 tacacctgca aaggcggcac aaccccagtg ccacgttgtg agttggatag ttgtggaaag
     2641 agtcaaatgg ctctcctcaa gcgtattcaa caaggggctg aaggatgccc agaaggtacc
     2701 ccattgtatg ggatctgatc tggggcctcg gtgcacatgc tttacatgtg tttagtcgag
     2761 gttaaaaaaa cgtctaggcc ccccgaacca cggggacgtg gttttccttt gaaaaacacg
     2821 atgataagct tgccacaacc cgggatcctc tagaatgtgt ggaaagaact gcttgctcat
     2881 agtcttcagc aaggaccagt tcccagaggt gtatgtgccc acagtgtttg agaactatgt
     2941 ggcagatatc gaggtggatg gaaagcaggt agagttggct ttgtgggaca cagctgggca
     3001 ggaagattat gatcgcctga ggcccctctc ctacccagat accgatgtta tactgatgtg
     3061 tttttccatc gacagccctg atagtttaga aaacatccca gaaaagtgga ccccagaagt
     3121 caagcatttc tgtcccaacg tgcccatcat cctggttggg aataagaagg atcttcggaa
     3181 tgatgagcac acaaggcggg agctagccaa gatgaagcag gagccggtga aacctgaaga
     3241 aggcagagat atggcaaaca ggattggcgc ttttgggtac atggagtgtt cagcaaagac
     3301 caaagatgga gtgagagagg tttttgaaat ggctacgaga gctgctctgc aagctgggga
     3361 tcccaattgg gagctcgtgt acacggcgcg cctgcaggga ggtggctcat ctggcggagg
     3421 tcagatctcg tacgcgtccc ggggcggagt gcaggtggaa accatctccc caggagacgg
     3481 gcgcaccttc cccaagcgcg gccagacctg cgtggtgcac tacaccggga tgcttgaaga
     3541 tggaaagaaa tttgattcct cccgggacag aaacaagccc tttaagttta tgctaggcaa
     3601 gcaggaggtg atccgaggct gggaagaagg ggttgcccag atgagtgtgg gtcagagagc
     3661 caaactgact atatctccag attatgccta tggtgccact gggcacccag gcatcatccc
     3721 accacatgcc actctcgtct tcgatgtgga gcttctaaaa ctggaaggcg gtggctcatc
     3781 tggcggaggt gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga
     3841 gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc
     3901 cacctacggc aagctgaccc tgaagctcat ctgcaccacc ggcaagctgc ccgtgccctg
     3961 gcccaccctc gtgaccaccc tcggctacgg cctgcagtgc ttcgcccgct accccgacca
     4021 catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac
     4081 catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga
     4141 caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct
     4201 ggggcacaag ctggagtaca actacaacag ccacaacgtc tatatcaccg ccgacaagca
     4261 gaagaacggc atcaaggcca acttcaagat ccgccacaac atcgaggacg gcggcgtgca
     4321 gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga
     4381 caaccactac ctgagctacc agtccaagct gagcaaagac cccaacgaga agcgcgatca
     4441 catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta
     4501 caagaaaaaa aagaagaaaa agagcaagac caaatgcgtg attatgtaag cggccgcttc
     4561 cctttagtga gggttaatgc ttcgagcaga catgataaga tacattgatg agtttggaca
     4621 aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc
     4681 tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt
     4741 tatgtttcag gttcaggggg agatgtggga ggttttttaa agcaagtaaa acctctacaa
     4801 atgtggtaaa atccgataag gatcgatccg ggctggcgta atagcgaaga ggcccgcacc
     4861 gatcgccctt cccaacagtt gcgcagcctg aatggcgaat ggacgcgccc tgtagcggcg
     4921 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc
     4981 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
     5041 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
     5101 accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
     5161 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
     5221 gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt
     5281 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa
     5341 tattaacgct tacaatttcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc
     5401 acaccgcata cgcggatctg cgcagcacca tggcctgaaa taacctctga aagaggaact
     5461 tggttaggta ccttctgagg cggaaagaac cagctgtgga atgtgtgtca gttagggtgt
     5521 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     5581 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     5641 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg
     5701 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
     5761 gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
     5821 ggcttttgca aaaagcttga ttcttctgac acaacagtct cgaacttaag gctagagcca
     5881 ccatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat
     5941 tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt
     6001 cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac
     6061 tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg
     6121 tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc
     6181 aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa
     6241 tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc
     6301 gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg
     6361 aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg
     6421 acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa
     6481 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg
     6541 acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct
     6601 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc
     6661 ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa
     6721 cctgccatca cgatggccgc aataaaatat ctttattttc attacatctg tgtgttggtt
     6781 ttttgtgtga atcgatagcg ataaggatcc gcgtatggtg cactctcagt acaatctgct
     6841 ctgatgccgc atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac
     6901 gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca
     6961 tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac
     7021 gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt
     7081 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     7141 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
     7201 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     7261 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac
     7321 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
     7381 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
     7441 gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
     7501 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
     7561 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
     7621 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
     7681 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
     7741 ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
     7801 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
     7861 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
     7921 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
     7981 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
     8041 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt
     8101 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     8161 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     8221 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     8281 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     8341 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
     8401 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     8461 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     8521 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     8581 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     8641 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     8701 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     8761 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     8821 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgg
     8881 ctcgacagat ct
//
