LOCUS       Exported                8480 bp ds-DNA     circular SYN 03-JUL-2023
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8480)
  AUTHORS   Triple Threat
  TITLE     Direct Submission
  JOURNAL   Exported Wednesday, Jul 5, 2023 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8480
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        295..572
                     /label=chicken beta-actin promoter
     misc_feature    573..801
                     /label=Hyrid Intron
     misc_feature    813..822
                     /label=Kozak sequence
     CDS             819..887
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="MDYKDHDGDYKDHDIDYKDDDDK"
     CDS             891..911
                     /codon_start=1
                     /product="nuclear localization signal of SV40 large T 
                     antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             912..2399
                     /codon_start=1
                     /label=enIscB
                     /translation="MMAVVYVISKSGKPLMPTTRCGHVRILLKEGKARVVERKPFTIQL
                     TYESAEETQPLVLGIDPGRTNIGMSVVTESGESVFNAQIRTRNKDVPKLMKDRKQYRMA
                     HRRLKRRCKRRRRAKAAGTAFEEGEKQRLLPGCFKPITCKSIRNKEARFNNRKRPVGWL
                     TPTANHLLVTHLNVVKKVQKILPVAKVVLELNRFSFMAMNNPKVQRWQYQRGPLYGKGS
                     VEEAVSMQQDGHCLFCKHGIDHYHHVVPRRKNGSETLENRVGLCEEHHRLVHTDKEWEA
                     NLASKKSGMNKKYHALSVLNQIIPYLADQLADMFPGNFCVTSGQDTYLFREEHGIPKDH
                     YLDAYCIACSALTDAKKVSSPKGRPYMVRQFRRHDRQACHKANLNRRYYMGGKLVATNR
                     HKAMDQKTDSLEEYRAAHSAADVSKLTVKHPSAQYKDMSRIMPGSILVSGEGKLFTLRR
                     SEGRNKGQVNYFVSTEGIKYWARKCQYLRNNGGLQIYV"
     CDS             2406..2504
                     /codon_start=1
                     /label=Linker
                     /translation="SGGSSGGSSGSETPGTSESATPESSGGSSGGSS"
     CDS             2505..3377
                     /codon_start=1
                     /label=T5E
                     /translation="MSKSWGKFIEEEEAEMASRRNLMIVDGTNLGFRFKHNNSKKPFAS
                     SYVSTIQSLAKSYSARTTIVLGDKGKSVFRLEHLPEYKGNRDEKYAQRTEEEKALDEQF
                     FEYLKDAFELCKTTFPTFTIRGVEADDMAAYIVKLIGHLYDHVWLISTDGDWDTLLTDK
                     VSRFSFTTRREYHLRDMYEHHNVDDVEQFISLKAIMGDLGDNIRGVEGIGAKRGYNIIR
                     EFGNVLDIIDQLPLPGKQKYIQNLNASEELLFRNLILVDLPTYCVDAIAAVGQDVLDKF
                     TKDILEIAEQ"
     CDS             3378..3425
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     polyA_signal    3459..3666
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     promoter        3667..3916
                     /label=U6 Promoter
                     /note="RNA polymerase III promoter for human U6 snRNA"
     enhancer        4206..4509
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4510..4713
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     CDS             4742..5452
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     rep_origin      6007..6462
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        6744..6848
                     /gene="bla"
                     /label=AmpR promoter
     CDS             6849..7709
                     /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"
     rep_origin      7880..8468
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 aggtacccgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
       61 gcccattgac gtcaatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt
      121 atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc
      181 ctattgacgt caatgacggt aaatggcccg cctggcattg tgcccagtac atgaccttat
      241 gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc atggtcgagg
      301 tgagccccac gttctgcttc actctcccca tctccccccc ctccccaccc ccaattttgt
      361 atttatttat tttttaatta ttttgtgcag cgatgggggc gggggggggg ggggggcgcg
      421 cgccaggcgg ggcggggcgg ggcgaggggc ggggcggggc gaggcggaga ggtgcggcgg
      481 cagccaatca gagcggcgcg ctccgaaagt ttccttttat ggcgaggcgg cggcggcggc
      541 ggccctataa aaagcgaagc gcgcggcggg cgggagtcgc tgcgacgctg ccttcgcccc
      601 gtgccccgct ccgccgccgc ctcgcgccgc ccgccccggc tctgactgac cgcgttactc
      661 ccacaggtga gcgggcggga cggcccttct cctccgggct gtaattagct gagcaagagg
      721 taagggttta agggatggtt ggttggtggg gtattaatgt ttaattacct ggagcacctg
      781 cctgaaatca ctttttttca ggttggaccg gtgccaccat ggactataag gaccacgacg
      841 gagactacaa ggatcatgat attgattaca aagacgatga cgataagatg cctaagaaga
      901 agagaaaggt gatgatggcc gtggtgtacg tgatcagcaa gtctggcaaa cccctgatgc
      961 ccaccacccg gtgcggccac gtgagaatcc tgctgaagga gggcaaggcc agagtggtgg
     1021 agcggaaacc gttcaccatc cagctgacct acgagagcgc ggaggaaaca cagcctctgg
     1081 tcctgggcat tgatcctgga cgcaccaata tcggcatgag cgtggtgaca gaatccggag
     1141 agagcgtgtt caacgcccag atcagaacca gaaacaagga cgtgcctaag ctgatgaaag
     1201 acagaaagca gtaccggatg gctcacagac ggctgaagcg gcgctgcaag cgaagaagaa
     1261 gagccaaggc cgccggcacc gcattcgagg aaggagaaaa gcagagactg ctgcctggct
     1321 gtttcaagcc tatcacatgt aagagcatca gaaataagga agctagattc aataacagga
     1381 agcggcctgt gggatggctg accccaaccg ccaaccacct gctggtgacc cacctgaacg
     1441 tggtcaagaa ggtgcaaaag atcctgcctg tggccaaggt ggtgctggaa ctgaatcggt
     1501 tcagcttcat ggccatgaac aaccccaagg tgcagcggtg gcagtaccag agaggccctc
     1561 tgtacggtaa aggcagcgtg gaagaggccg tgagcatgca gcaggacggc cactgcctgt
     1621 tttgcaagca cggcattgac cactatcacc atgtggttcc taggagaaag aacggctctg
     1681 aaaccctgga gaaccgggtg ggcctgtgcg aggaacacca ccggctcgtc cacacagata
     1741 aggagtggga ggccaatctg gcttctaaga agagcggaat gaacaagaaa taccacgccc
     1801 tgagcgtgct gaaccagatc atcccatacc tggccgacca gcttgccgat atgttccccg
     1861 gcaacttctg tgtgaccagc ggacaggata catacctgtt tcgggaagaa catggcatcc
     1921 ccaaggacca ctacctggat gcctactgca tcgcctgcag cgccctgacc gacgccaaga
     1981 aggtgagcag ccctaaaggc agaccttaca tggtgagaca gttcagacgc cacgatagac
     2041 aggcctgtca caaggccaac ctgaacagaa gatactacat gggcggcaaa ctggttgcta
     2101 caaacagaca taaggccatg gaccagaaaa ccgacagcct ggaggagtac agagccgccc
     2161 acagcgccgc tgacgtatcc aagctgacag tgaagcaccc ttctgctcaa tataaggaca
     2221 tgtcccggat catgcctggc agcatcctcg tgtccggcga gggcaagctc tttacactga
     2281 gaagatccga gggcagaaac aagggccaag tgaactactt cgtgagcacc gagggcatca
     2341 agtactgggc cagaaagtgc cagtatctga gaaacaatgg cggcctgcag atctacgtga
     2401 agctttccgg aggatctagc ggaggctcct ctggcagcga gacaccagga acaagcgagt
     2461 cagcaacacc agagagcagt ggcggcagca gcggcggcag cagtatgagc aagtcctggg
     2521 gcaagttcat cgaggaagag gaagctgaaa tggcctctag acgcaacctg atgatcgtgg
     2581 atggcaccaa cctcggcttc aggttcaagc acaacaactc caagaaacct ttcgccagca
     2641 gctacgtgtc taccatccag agcctggcca agagctatag cgccagaacc acaatcgtgc
     2701 tcggcgataa gggcaaaagc gtgttcagac tggaacatct gcctgagtac aagggcaacc
     2761 gggacgagaa gtacgcccag cggaccgagg aagagaaggc tctggatgag cagtttttcg
     2821 agtacctgaa agacgccttc gagctttgta aaacaacatt tcctacattc acaatcagag
     2881 gagtggaagc cgatgatatg gccgcttata tcgtgaagct gatcggccac ctgtacgacc
     2941 acgtgtggct gatctctaca gatggtgact gggacaccct gctgaccgac aaggtgtcca
     3001 gattcagctt caccaccaga agagagtacc acctgagaga tatgtacgag caccacaatg
     3061 tggacgacgt ggagcaattt atcagcctga aggccattat gggcgacctg ggcgacaaca
     3121 tccggggcgt tgaaggcatc ggcgctaaaa gaggctacaa catcatccgg gagttcggca
     3181 atgtgctgga catcatcgac cagctgcccc tgccaggcaa gcagaaatac atccagaacc
     3241 tgaacgccag cgaggagctg ctgttcagaa atctgattct ggtggacctg cctacctact
     3301 gcgtggatgc catcgccgcc gtgggccagg acgtgctgga caagtttacc aaggacatcc
     3361 tggaaatcgc cgagcagaaa aggccggcgg ccacgaagaa ggccggccag gcaaagaaga
     3421 aaaagtaaga attcctagag ctcgctgatc agcctcgact gtgccttcta gttgccagcc
     3481 atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt
     3541 cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct
     3601 ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagagaata gcaggcatgc
     3661 tggggagagg gcctatttcc catgattcct tcatatttgc atatacgata caaggctgtt
     3721 agagagataa ttggaattaa tttgactgta aacacaaaga tattagtaca aaatacgtga
     3781 cgtagaaagt aataatttct tgggtagttt gcagttttaa aattatgttt taaaatggac
     3841 tatcatatgc ttaccgtaac ttgaaagtat ttcgatttct tggctttata tatcttgtgg
     3901 aaaggacgaa acaccgggag tcttccgaag acttggctcg tccaactgcg gttgaacgag
     3961 cacaggctga gacattcgta aggccgaaag gccggacgca ccctgggatt tccccagtcc
     4021 ccggaactgc atagcggatg ccagttgatg gagcaatcta tcagataagc cagggggaac
     4081 aatcacctct ctgtatcaga gagagtttta caaaaggagg aacggtttta tttttttgcg
     4141 gccgctagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
     4201 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
     4261 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
     4321 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
     4381 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
     4441 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
     4501 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
     4561 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
     4621 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
     4681 cgtgtacggt gggaggtcta tataagcaga gctggtttag tgaaccgtca gatccgctag
     4741 catggtgagc aagggcgagg aggataacat ggccatcatc aaggagttca tgcgcttcaa
     4801 ggtgcacatg gagggctccg tgaacggcca cgagttcgag atcgagggcg agggcgaggg
     4861 ccgcccctac gagggcaccc agaccgccaa gctgaaggtg accaagggtg gccccctgcc
     4921 cttcgcctgg gacatcctgt cccctcagtt catgtacggc tccaaggcct acgtgaagca
     4981 ccccgccgac atccccgact acttgaagct gtccttcccc gagggcttca agtgggagcg
     5041 cgtgatgaac ttcgaggacg gcggcgtggt gaccgtgacc caggactcct ccctgcagga
     5101 cggcgagttc atctacaagg tgaagctgcg cggcaccaac ttcccctccg acggccccgt
     5161 aatgcagaag aaaaccatgg gctgggaggc ctcctccgag cggatgtacc ccgaggacgg
     5221 cgccctgaag ggcgagatca agcagaggct gaagctgaag gacggcggcc actacgacgc
     5281 tgaggtcaag accacctaca aggccaagaa gcccgtgcag ctgcccggcg cctacaacgt
     5341 caacatcaag ttggacatca cctcccacaa cgaggactac accatcgtgg aacagtacga
     5401 acgcgccgag ggccgccact ccaccggcgg catggacgag ctgtacaagt aaggatccgg
     5461 gatgcagaaa ttgatgatct attaaacaat aaagatgtcc actaaaatgg aagtttttcc
     5521 tgtcatactt tgttaagaag ggtgagaaca gagtacctac attttgaatg gaaggattgg
     5581 agctacgggg gtgggggtgg ggtgggatta gataaatgcc tgctctttac tgaaggctct
     5641 ttactattgc tttatgataa tgtttcatag ttggatatca taatttaaac aagcaaaacc
     5701 aaattaaggg ccagctcatt cctcccactc atgatctata gatctataga tctctcgtgg
     5761 gatcattgtt tttctcttga ttcccacttt gtggttctaa gtactgtggt ttccaaatgt
     5821 gtcagtttca tagcctgaag aacgagatca gcagcctctg ttccacatac acttcattct
     5881 cagtattgtt ttgccaagtt ctaattccat cagaagctgg tcgacctgca ggggcgcctg
     5941 atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatacg tcaaagcaac
     6001 catagtacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
     6061 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc
     6121 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
     6181 gatttagtgc tttacggcac ctcgacccca aaaaacttga tttgggtgat ggttcacgta
     6241 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
     6301 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcgggc tattcttttg
     6361 atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa
     6421 aatttaacgc gaattttaac aaaatattaa cgtttacaat tttatggtgc actctcagta
     6481 caatctgctc tgatgccgca tagttaagcc agccccgaca cccgccaaca cccgctgacg
     6541 cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg
     6601 ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc
     6661 tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag
     6721 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     6781 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     6841 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
     6901 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
     6961 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
     7021 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
     7081 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
     7141 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
     7201 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
     7261 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
     7321 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
     7381 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
     7441 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
     7501 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
     7561 gtggaagccg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
     7621 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
     7681 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
     7741 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     7801 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     7861 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     7921 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     7981 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc
     8041 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     8101 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     8161 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     8221 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     8281 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     8341 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     8401 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     8461 tatggaaaaa cgccagcaac
//
