LOCUS       Exported                9624 bp ds-DNA     circular SYN 15-JUL-2023
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9624)
  AUTHORS   gene
  TITLE     Direct Submission
  JOURNAL   Exported Friday, Dec 20, 2024 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..9624
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     source          1091..2866
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     source          2867..2956
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     source          3614..4132
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     source          4133..4222
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     source          4223..5281
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     misc_feature    1..659
                     /label=CMV IE Enhancer
     misc_feature    669..750
                     /label=CMV IE Promoter
     misc_feature    890..1022
                     /label=IVS
     misc_feature    1067..1085
                     /label=T7 RNA Polymerase Promoter
     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"
     misc_feature    1832..2866
                     /label=GID1
     misc_feature    2853..2878
                     /label=overlap region
     misc_feature    2918..2953
                     /label=Cdc42-N12
     misc_feature    2991..3586
                     /label=IRES
     misc_feature    3608..3613
                     /label=Must do C13 part first, can use XbaI
     misc_feature    3614..3616
                     /label=Cdc42-13C (no caax)
     misc_feature    3620..4132
                     /label=Cdc42-C13 (T17N)
     misc_feature    4136..4155
                     /label=overlap region
     misc_feature    4223..4498
                     /label=GAI (1-92)
     CDS             4235..4252
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             4523..5236
                     /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"
     misc_feature    complement(5290..5311)
                     /label=T3 RNA Polymerase Promoter
     misc_feature    5321..5542
                     /label=SV40 polyA signal
     misc_feature    5637..6091
                     /label=f1 ori
     misc_feature    6154..6571
                     /label=SV40 Enhancer/Early Promoter
     misc_feature    6470..6535
                     /label=SV40 ori
     misc_feature    6615..7409
                     /label=Neo (R)
     misc_feature    7932..8792
                     /label=Amp (R)
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 tatggctgcg agcgatgaag ttaatcttat
     1861 tgagagcaga acagtggttc ctctcaatac atgggtttta atatccaact tcaaagtagc
     1921 ctacaatatc cttcgtcgcc ctgatggaac ctttaaccga cacttagctg agtatctaga
     1981 ccgtaaagtc actgcaaacg ccaatccggt tgatggggtt ttctcgttcg atgtcttgat
     2041 tgatcgcagg atcaatcttc taagcagagt ctatagacca gcttatgcag atcaagagca
     2101 acctcctagt attttagatc tcgagaagcc tgttgatggc gacattgtcc ctgttatatt
     2161 gttcttccat ggaggtagct ttgctcattc ttctgcaaac agtgccatct acgatactct
     2221 ttgtcgcagg cttgttggtt tgtgcaagtg tgttgttgtc tctgtgaatt atcggcgtgc
     2281 accagagaat ccataccctt gtgcttatga tgatggttgg attgctctta attgggttaa
     2341 ctcgagatct tggcttaaat ccaagaaaga ctcaaaggtc catattttct tggctggtga
     2401 tagctctgga ggtaacatcg cgcataatgt ggctttaaga gcgggtgaat cgggaatcga
     2461 tgttttgggg aacattctgc tgaatcctat gtttggtggg aatgagagaa cggagtctga
     2521 gaaaagtttg gatgggaaat actttgtgac ggttagagac cgcgattggt actggaaagc
     2581 gtttttaccc gagggagaag atagagagca tccagcgtgt aatccgttta gcccgagagg
     2641 gaaaagctta gaaggagtga gtttccccaa gagtcttgtg gttgtcgcgg gtttggattt
     2701 gattagagat tggcagttgg catacgcgga agggctcaag aaagcgggtc aagaggttaa
     2761 gcttatgcat ttagagaaag caactgttgg gttttacctc ttgcctaata acaatcattt
     2821 ccataatgtt atggatgaga tttcggcgtt tgtaaacgcg gaatgtcaga tctcgtacgc
     2881 gtcccggggc ggtggctcat ctggcggagg tgagctccag acaattaagt gtgttgttgt
     2941 gggcgatggt gcttaagaat tcacgcgtcg agcatgcatc tagggcggcc aattccgccc
     3001 ctctcccccc cccccctctc cctccccccc ccctaacgtt actggccgaa gccgcttgga
     3061 ataaggccgg tgtgcgtttg tctatatgtt attttccacc atattgccgt cttttggcaa
     3121 tgtgagggcc cggaaacctg gccctgtctt cttgacgagc attcctaggg gtctttcccc
     3181 tctcgccaaa ggaatgcaag gtctgttgaa tgtcgtgaag gaagcagttc ctctggaagc
     3241 ttcttgaaga caaacaacgt ctgtagcgac cctttgcagg cagcggaacc ccccacctgg
     3301 cgacaggtgc ctctgcggcc aaaagccacg tgtataagat acacctgcaa aggcggcaca
     3361 accccagtgc cacgttgtga gttggatagt tgtggaaaga gtcaaatggc tctcctcaag
     3421 cgtattcaac aaggggctga aggatgccca gaaggtaccc cattgtatgg gatctgatct
     3481 ggggcctcgg tgcacatgct ttacatgtgt ttagtcgagg ttaaaaaaac gtctaggccc
     3541 cccgaaccac ggggacgtgg ttttcctttg aaaaacacga tgataagctt gccacaaccc
     3601 gggatcctct agaatggggg ttggtaaaaa ctgtctcctg atatcctaca caacaaacaa
     3661 atttccatcg gagtatgtac cgactgtttt tgacaactat gcagtcacag ttatgattgg
     3721 tggagaacca tatactcttg gactttttga tactgcaggg caagaggatt atgacagatt
     3781 acgaccgctg agttatccac aaacagatgt atttctagtc tgtttttcag tggtctctcc
     3841 atcttcattt gaaaacgtga aagaaaagtg ggtgcctgag ataactcacc actgtccaaa
     3901 gactcctttc ttgcttgttg ggactcaaat tgatctcaga gatgacccct ctactattga
     3961 gaaacttgcc aagaacaaac agaagcctat cactccagag actgctgaaa agctggcccg
     4021 tgacctgaag gctgtcaagt atgtggagtg ttctgcactt acacagagag gtctgaagaa
     4081 tgtgtttgat gaggctatcc tagctgccct cgagcctccg gaaactcaac ccggtgatcc
     4141 caattgggag ctcgtgtaca cggcgcgcct gcagggaggt ggctcatctg gcggaggtca
     4201 gatctcgtac gcgtcccggg gcatgaagag agatcatcat catcatcatc atcaagataa
     4261 gaagactatg atgatgaatg aagaagacga cggtaacggc atggatgagc ttctagctgt
     4321 tcttggttac aaggttaggt catccgaaat ggctgatgtt gctcagaaac tcgagcagct
     4381 tgaagttatg atgtctaatg ttcaagaaga cgatctttct caactcgcta ctgagactgt
     4441 tcactataat ccggcggagc tttacacgtg gcttgattct atgctcaccg accttaatgg
     4501 cggtggctca tctggcggag gtgtgagcaa gggcgaggag ctgttcaccg gggtggtgcc
     4561 catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg
     4621 cgagggcgat gccacctacg gcaagctgac cctgaagctc atctgcacca ccggcaagct
     4681 gcccgtgccc tggcccaccc tcgtgaccac cctcggctac ggcctgcagt gcttcgcccg
     4741 ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt
     4801 ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa
     4861 gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga
     4921 cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg tctatatcac
     4981 cgccgacaag cagaagaacg gcatcaaggc caacttcaag atccgccaca acatcgagga
     5041 cggcggcgtg cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt
     5101 gctgctgccc gacaaccact acctgagcta ccagtccaag ctgagcaaag accccaacga
     5161 gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat
     5221 ggacgagctg tacaagaaaa aaaagaagaa aaagagcaag accaaatgcg tgattatgta
     5281 agcggccgct tccctttagt gagggttaat gcttcgagca gacatgataa gatacattga
     5341 tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg
     5401 tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagtta acaacaacaa
     5461 ttgcattcat tttatgtttc aggttcaggg ggagatgtgg gaggtttttt aaagcaagta
     5521 aaacctctac aaatgtggta aaatccgata aggatcgatc cgggctggcg taatagcgaa
     5581 gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga atggacgcgc
     5641 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     5701 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     5761 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
     5821 tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc
     5881 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
     5941 tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga
     6001 ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
     6061 attttaacaa aatattaacg cttacaattt cctgatgcgg tattttctcc ttacgcatct
     6121 gtgcggtatt tcacaccgca tacgcggatc tgcgcagcac catggcctga aataacctct
     6181 gaaagaggaa cttggttagg taccttctga ggcggaaaga accagctgtg gaatgtgtgt
     6241 cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     6301 ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     6361 caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg
     6421 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
     6481 tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt
     6541 tttggaggcc taggcttttg caaaaagctt gattcttctg acacaacagt ctcgaactta
     6601 aggctagagc caccatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg
     6661 tggagaggct attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg
     6721 tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg
     6781 ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc
     6841 cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg
     6901 aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca
     6961 tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc
     7021 aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg
     7081 atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg
     7141 cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata
     7201 tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg
     7261 accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat
     7321 gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct
     7381 tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca
     7441 agcgacgccc aacctgccat cacgatggcc gcaataaaat atctttattt tcattacatc
     7501 tgtgtgttgg ttttttgtgt gaatcgatag cgataaggat ccgcgtatgg tgcactctca
     7561 gtacaatctg ctctgatgcc gcatagttaa gccagccccg acacccgcca acacccgctg
     7621 acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct
     7681 ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg
     7741 gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt
     7801 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     7861 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     7921 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     7981 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     8041 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     8101 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
     8161 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     8221 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
     8281 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     8341 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     8401 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     8461 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
     8521 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
     8581 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     8641 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     8701 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     8761 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
     8821 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     8881 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     8941 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     9001 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     9061 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     9121 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     9181 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     9241 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     9301 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     9361 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     9421 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     9481 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     9541 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     9601 ttgctcacat ggctcgacag atct
//
