LOCUS       Exported                8242 bp DNA     circular SYN 07-JUN-2022
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8242)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jun 27, 2022 from SnapGene Viewer 6.0.6
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8242
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        8..387
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        83..1678
                     /label=CAG promoter
     promoter        390..665
                     /label=chicken beta-actin promoter
     promoter        1739..1755
                     /label=T7 promoter
     misc_feature    1757..1765
                     /label=Kozak sequence
     CDS             1769..1789
                     /codon_start=1
                     /label=NLS
                     /translation="PKKKRKV"
     CDS             1796..4729
                     /codon_start=1
                     /label=PlmCasX
                     /translation="MQEIKRINKIRRRLVKDSNTKKAGKTGPMKTLLVRVMTPDLRERL
                     ENLRKKPENIPQPISNTSRANLNKLLTDYTEMKKAILHVYWEEFQKDPVGLMSRVAQPA
                     PKNIDQRKLIPVKDGNERLTSSGFACSQCCQPLYVYKLEQVNDKGKPHTNYFGRCNVSE
                     HERLILLSPHKPEANDELVTYSLGKFGQRALDFYSIHVTRESNHPVKPLEQIGGNSCAS
                     GPVGKALSDACMGAVASFLTKYQDIILEHQKVIKKNEKRLANLKDIASANGLAFPKITL
                     PPQPHTKEGIEAYNNVVAQIVIWVNLNLWQKLKIGRDEAKPLQRLKGFPSFPLVERQAN
                     EVDWWDMVCNVKKLINEKKEDGKVFWQNLAGYKRQEALLPYLSSEEDRKKGKKFARYQF
                     GDLLLHLEKKHGEDWGKVYDEAWERIDKKVEGLSKHIKLEEERRSEDAQSKAALTDWLR
                     AKASFVIEGLKEADKDEFCRCELKLQKWYGDLRGKPFAIEAENSILDISGFSKQYNCAF
                     IWQKDGVKKLNLYLIINYFKGGKLRFKKIKPEAFEANRFYTVINKKSGEIVPMEVNFNF
                     DDPNLIILPLAFGKRQGREFIWNDLLSLETGSLKLANGRVIEKTLYNRRTRQDEPALFV
                     ALTFERREVLDSSNIKPMNLIGIDRGENIPAVIALTDPEGCPLSRFKDSLGNPTHILRI
                     GESYKEKQRTIQAAKEVEQRRAGGYSRKYASKAKNLADDMVRNTARDLLYYAVTQDAML
                     IFENLSRGFGRQGKRTFMAERQYTRMEDWLTAKLAYEGLPSKTYLSKTLAQYTSKTCSN
                     CGFTITSADYDRVLEKLKKTATGWMTTINGKELKVEGQITYYNRYKRQNVVKDLSVELD
                     RLSEESVNNDISSWTKGRSGEALSLLKKRFSHRPVQEKFVCLNCGFETHADEQAALNIA
                     RSWLFLRSQEYKKYQTNKTTGNTDKRAFVETWQSFYRKKLKEVWKPAV"
     CDS             4736..4756
                     /codon_start=1
                     /label=NLS
                     /translation="PKKKRKV"
     CDS             4763..4852
                     /codon_start=1
                     /label=3xHA tag
                     /translation="YPYDVPDYAGYPYDVPDYAGSYPYDVPDYA"
     CDS             4874..4927
                     /codon_start=1
                     /label=T2A peptide
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             4928..5644
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4991..5010)
                     /label=S-GFP-1R
     polyA_signal    5690..5811
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5854..5870)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    5878..5894
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(5902..5932)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     rep_origin      complement(6256..6844)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number colE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(7015..7875)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(7876..7980)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 acgcgttgac attgattatt gactagttat taatagtaat caattacggg gtcattagtt
       61 catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga
      121 ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca
      181 atagggactt tccattgacg tcaatgggtg gactatttac ggtaaactgc ccacttggca
      241 gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg
      301 cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc
      361 tacgtattag tcatcgctat taccatgggt cgaggtgagc cccacgttct gcttcactct
      421 ccccatctcc cccccctccc cacccccaat tttgtattta tttatttttt aattattttg
      481 tgcagcgatg ggggcggggg gggggggggc gcgcgccagg cggggcgggg cggggcgagg
      541 ggcggggcgg ggcgaggcgg agaggtgcgg cggcagccaa tcagagcggc gcgctccgaa
      601 agtttccttt tatggcgagg cggcggcggc ggcggcccta taaaaagcga agcgcgcggc
      661 gggcgggagt cgctgcgttg ccttcgcccc gtgccccgct ccgcgccgcc tcgcgccgcc
      721 cgccccggct ctgactgacc gcgttactcc cacaggtgag cgggcgggac ggcccttctc
      781 ctccgggctg taattagcgc ttggtttaat gacggctcgt ttcttttctg tggctgcgtg
      841 aaagccttaa agggctccgg gagggccctt tgtgcggggg ggagcggctc ggggggtgcg
      901 tgcgtgtgtg tgtgcgtggg gagcgccgcg tgcggcccgc gctgcccggc ggctgtgagc
      961 gctgcgggcg cggcgcgggg ctttgtgcgc tccgcgtgtg cgcgagggga gcgcggccgg
     1021 gggcggtgcc ccgcggtgcg ggggggctgc gaggggaaca aaggctgcgt gcggggtgtg
     1081 tgcgtggggg ggtgagcagg gggtgtgggc gcggcggtcg ggctgtaacc cccccctgca
     1141 cccccctccc cgagttgctg agcacggccc ggcttcgggt gcggggctcc gtacggggcg
     1201 tggcgcgggg ctcgccgtgc cgggcggggg gtggcggcag gtgggggtgc cgggcggggc
     1261 ggggccgcct cgggccgggg agggctcggg ggaggggcgc ggcggccccc ggagcgccgg
     1321 cggctgtcga ggcgcggcga gccgcagcca ttgcctttta tggtaatcgt gcgagagggc
     1381 gcagggactt cctttgtccc aaatctgtgc ggagccgaaa tctgggaggc gccgccgcac
     1441 cccctctagc gggcgcgggg cgaagcggtg cggcgccggc aggaaggaaa tgggcgggga
     1501 gggccttcgt gcgtcgccgc gccgccgtcc ccttctccat ctccagcctc ggggctgtcc
     1561 gcagggggac ggctgccttc gggggggacg gggcagggcg gggttcggct tctggcgtgt
     1621 gaccggcggc tctagcgcct ctgctaacca tgttcatgcc ttcttctttt tcctacagct
     1681 cctgggcaac gtgctggtta ttgtgctgtc tcatcatttt ggcaaagcta gtgaattcta
     1741 atacgactca ctataggccg ccaccatgcc caagaagaag aggaaggttc ccgggatgca
     1801 ggagatcaag aggatcaata agatccggag aaggctggtg aaggacagca acaccaagaa
     1861 ggccggcaag accggcccca tgaagacact gctggtgaga gtgatgacac ctgatctgcg
     1921 cgagcggctg gagaacctga gaaagaagcc tgagaatatc ccacagccca tctctaacac
     1981 cagccgggcc aacctgaata agctgctgac cgactataca gagatgaaga aggccatcct
     2041 gcacgtgtac tgggaggagt tccagaagga ccccgtgggc ctgatgtcca gagtggccca
     2101 gcctgcccca aagaatatcg accagcggaa gctgatcccc gtgaaggatg gcaacgagag
     2161 actgaccagc tccggcttcg cctgctctca gtgctgtcag cctctgtacg tgtataagct
     2221 ggagcaagtg aatgacaagg gcaagccaca cacaaactac tttggccggt gtaacgtgag
     2281 cgagcacgag agactgatcc tgctgtctcc ccacaagcct gaggccaacg acgagctggt
     2341 gacctattcc ctgggcaagt tcggacagcg ggccctggac ttctactcta tccacgtgac
     2401 acgcgagagc aatcacccag tgaagcccct ggagcagatc ggaggcaact cctgcgcctc
     2461 tggaccagtg ggcaaggccc tgagcgacgc atgtatggga gcagtggcct ccttcctgac
     2521 caagtatcag gatatcatcc tggagcacca gaaagtgatc aagaagaacg agaagcggct
     2581 ggccaatctg aaggacatcg cctccgccaa cggcctggcc tttcctaaga tcaccctgcc
     2641 ccctcagcca cacacaaagg agggcatcga ggcctacaac aatgtggtgg cccagatcgt
     2701 gatctgggtg aacctgaatc tgtggcagaa gctgaagatc ggaagggacg aggcaaagcc
     2761 actgcagcgc ctgaagggct tcccttcttt tccactggtg gagaggcagg ccaatgaggt
     2821 ggactggtgg gatatggtgt gcaacgtgaa gaagctgatc aatgagaaga aggaggatgg
     2881 caaggtgttc tggcagaacc tggccggcta taagcgccag gaggccctgc tgccttacct
     2941 gtctagcgag gaggaccgga agaagggcaa gaagttcgcc agatatcagt ttggcgatct
     3001 gctgctgcac ctggagaaga agcacggcga ggactggggc aaggtgtacg atgaggcctg
     3061 ggagaggatc gacaagaagg tggagggcct gtctaagcac atcaagctgg aggaggagcg
     3121 ccggagcgag gacgcccagt ccaaggccgc cctgaccgat tggctgaggg ccaaggccag
     3181 cttcgtgatc gagggcctga aggaggccga caaggatgag ttttgccggt gtgagctgaa
     3241 gctgcagaag tggtatggcg acctgagagg caagcccttt gccatcgagg ccgagaatag
     3301 catcctggat atcagcggct tctccaagca gtacaactgc gcctttatct ggcagaagga
     3361 tggcgtgaag aagctgaacc tgtacctgat catcaattat ttcaagggcg gcaagctgag
     3421 gtttaagaag atcaagcccg aggccttcga ggccaatcgc ttttatacag tgatcaacaa
     3481 gaagtccggc gagatcgtgc ctatggaggt gaacttcaat tttgacgatc caaatctgat
     3541 catcctgccc ctggccttcg gcaagagaca gggcagggag tttatctgga acgacctgct
     3601 gagcctggag acaggcagcc tgaagctggc caatggcaga gtgatcgaga agaccctgta
     3661 caacagaagg acacgccagg atgagcctgc cctgttcgtg gccctgacat ttgagcgccg
     3721 ggaggtgctg gactcctcta acatcaagcc aatgaatctg atcggcatcg atcggggcga
     3781 gaatatccca gccgtgatcg ccctgaccga ccccgaggga tgtcctctgt ccagattcaa
     3841 ggattctctg ggcaacccaa cccacatcct gaggatcggc gagagctata aggagaagca
     3901 gcgcacaatc caggccgcca aggaggtgga gcagagaagg gcaggaggat acagccggaa
     3961 gtatgcctcc aaggccaaga atctggccga cgatatggtg agaaacaccg ccagggacct
     4021 gctgtactat gccgtgacac aggatgccat gctgatcttc gagaacctgt ctcgcggctt
     4081 tggccggcag ggcaagagaa ccttcatggc cgagaggcag tacacccgca tggaggactg
     4141 gctgacagcc aagctggcct acgagggcct gccttctaag acctatctga gcaagacact
     4201 ggcccagtac acctccaaga catgctctaa ttgtggcttt accatcacaa gcgccgacta
     4261 tgatagagtg ctggagaagc tgaagaagac cgccacaggc tggatgacca caatcaacgg
     4321 caaggagctg aaggtggagg gccagatcac atactataac cgctacaagc ggcagaatgt
     4381 ggtgaaggac ctgagcgtgg agctggatag gctgtctgag gagagcgtga acaatgatat
     4441 cagctcctgg accaagggcc gctccggaga ggccctgtct ctgctgaaga agcggttctc
     4501 ccacagacca gtgcaggaga agttcgtgtg cctgaattgt ggctttgaga cacacgcaga
     4561 cgagcaggcc gccctgaaca tcgccaggag ctggctgttt ctgcgctccc aggagtacaa
     4621 gaagtatcag accaacaaga ccacaggcaa tacagataag cgggccttcg tggagacatg
     4681 gcagtccttt tacagaaaga agctgaagga agtgtggaag cccgccgtgg ctagcccaaa
     4741 gaagaagagg aaagtctcta gataccctta tgatgttcca gattatgccg gatacccata
     4801 cgatgtccct gactatgcag gctcctaccc ttatgacgtc ccagactacg ccggatccag
     4861 gtccggcggc ggagagggca gaggaagtct tctaacatgc ggtgacgtgg aggagaatcc
     4921 cggcccaatg gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga
     4981 gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc
     5041 cacctacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg
     5101 gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct accccgacca
     5161 catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac
     5221 catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga
     5281 caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct
     5341 ggggcacaag ctggagtaca actacaacag ccacaacgtc tatatcatgg ccgacaagca
     5401 gaagaacggc atcaaggtga acttcaagat ccgccacaac atcgaggacg gcagcgtgca
     5461 gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga
     5521 caaccactac ctgagcaccc agtccgccct gagcaaagac cccaacgaga agcgcgatca
     5581 catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta
     5641 caagtaactg cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat
     5701 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     5761 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt attaattaac
     5821 gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
     5881 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctaggg
     5941 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     6001 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     6061 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     6121 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     6181 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     6241 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     6301 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     6361 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     6421 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     6481 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     6541 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     6601 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     6661 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     6721 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     6781 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     6841 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     6901 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     6961 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     7021 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     7081 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     7141 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     7201 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     7261 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     7321 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     7381 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     7441 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     7501 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     7561 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     7621 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     7681 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     7741 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     7801 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     7861 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     7921 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     7981 cacatttccc cgaaaagtgc cacctgacgt cgacggatcg ggagatcgat ctcccgatcc
     8041 cctagggtcg actctcagta caatctgctc tgatgccgca tagttaagcc agtatctgct
     8101 ccctgcttgt gtgttggagg tcgctgagta gtgcgcgagc aaaatttaag ctacaacaag
     8161 gcaaggcttg accgacaatt gcatgaagaa tctgcttagg gttaggcgtt ttgcgctgct
     8221 tcgcgatgta cgggccagat at
//
