LOCUS       Exported                8723 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 8723)
  AUTHORS   qqxiao
  TITLE     Direct Submission
  JOURNAL   Exported Wednesday, Jul 5, 2023 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8723
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     rep_origin      30..618
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     promoter        686..926
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA"
     misc_feature    1010
                     /label=A-to-G R2M4
     enhancer        1158..1443
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     promoter        1445..1722
                     /label=chicken beta-actin promoter
     intron          1723..1951
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     CDS             1972..2025
                     /codon_start=1
                     /label=bp SV40 NLS
                     /translation="KRTADGSEFESPKKKRKV"
     CDS             2026..2523
                     /codon_start=1
                     /label=TAD8e (V106W)
                     /translation="SEVEFSHEYWMRHALTLAKRARDEREVPVGAVLVLNNRVIGEGWN
                     RAIGLHDPTAHAEIMALRQGGLVMQNYRLIDATLYVTFEPCVMCAGAMIHSRIGRVVFG
                     WRNSKRGAAGSLMNVLNYPGMNHRVEITEGILADECAALLCDFYRMPRQVFNAQKKAQS
                     SIN"
     misc_feature    2338..2340
                     /label=V106W
     CDS             2524..2619
                     /codon_start=1
                     /label=LINKER
                     /translation="SGGSSGGSSGSETPGTSESATPESSGGSSGGS"
     CDS             2620..4107
                     /codon_start=1
                     /label=enIscB
                     /translation="MMAVVYVISKSGKPLMPTTRCGHVRILLKEGKARVVERKPFTIQL
                     TYESAEETQPLVLGIAPGRTNIGMSVVTESGESVFNAQIRTRNKDVPKLMKDRKQYRMA
                     HRRLKRRCKRRRRAKAAGTAFEEGEKQRLLPGCFKPITCKSIRNKEARFNNRKRPVGWL
                     TPTANHLLVTHLNVVKKVQKILPVAKVVLELNRFSFMAMNNPKVQRWQYQRGPLYGKGS
                     VEEAVSMQQDGHCLFCKHGIDHYHHVVPRRKNGSETLENRVGLCEEHHRLVHTDKEWEA
                     NLASKKSGMNKKYHALSVLNQIIPYLADQLADMFPGNFCVTSGQDTYLFREEHGIPKDH
                     YLDAYCIACSALTDAKKVSSPKGRPYMVRQFRRHDRQACHKANLNRRYYMGGKLVATNR
                     HKAMDQKTDSLEEYRAAHSAADVSKLTVKHPSAQYKDMSRIMPGSILVSGEGKLFTLRR
                     SEGRNKGQVNYFVSTEGIKYWARKCQYLRNNGGLQIYV"
     misc_feature    2800..2802
                     /label=D61A
     CDS             4132..4185
                     /codon_start=1
                     /label=bpSV40 NLS
                     /translation="KRTADGSEFESPKKKRKV"
     CDS             4210..4707
                     /codon_start=1
                     /label=TAD8e (V106W)
                     /translation="SEVEFSHEYWMRHALTLAKRARDEREVPVGAVLVLNNRVIGEGWN
                     RAIGLHDPTAHAEIMALRQGGLVMQNYRLIDATLYVTFEPCVMCAGAMIHSRIGRVVFG
                     WRNSKRGAAGSLMNVLNYPGMNHRVEITEGILADECAALLCDFYRMPRQVFNAQKKAQS
                     SIN"
     misc_feature    4522..4524
                     /label=V106W
     CDS             4708..4761
                     /codon_start=1
                     /label=bpNLS
                     /translation="KRTADGSEFESPKKKRKV"
     polyA_signal    4795..5002
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     enhancer        5071..5374
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5375..5578
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     misc_feature    5607..5615
                     /label=Kozak
     CDS             5613..6323
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     DDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     rep_origin      6880..7335
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        7617..7721
                     /gene="bla"
                     /label=AmpR promoter
     CDS             7722..8582
                     /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"
ORIGIN
        1 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
       61 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
      121 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt
      181 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
      241 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
      301 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
      361 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
      421 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
      481 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
      541 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
      601 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
      661 tggccttttg ctcacatgct ctagagaggg cctatttccc atgattcctt catatttgca
      721 tatacgatac aaggctgtta gagagataat tggaattaat ttgactgtaa acacaaagat
      781 attagtacaa aatacgtgac gtagaaagta ataatttctt gggtagtttg cagttttaaa
      841 attatgtttt aaaatggact atcatatgct taccgtaact tgaaagtatt tcgatttctt
      901 ggctttatat atcttgtgga aaggacgaaa caccggggtc ttcgagaaga cctggctcgt
      961 ccaactgcgg ttgaacgagc acaggctgag acattcgtaa ggccgaaagg ccggacgcac
     1021 cctgggattt ccccagtccc cggaactgca tagcggatgc cagttgatgg agcaatctat
     1081 cagataagcc agggggaaca atcacctctc tgtatcagag agagttttac aaaaggagga
     1141 acggtttttt tggtacccgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc
     1201 aacgaccccc gcccattgac gtcaatagta acgccaatag ggactttcca ttgacgtcaa
     1261 tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca
     1321 agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcattg tgcccagtac
     1381 atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc
     1441 atggtcgagg tgagccccac gttctgcttc actctcccca tctccccccc ctccccaccc
     1501 ccaattttgt atttatttat tttttaatta ttttgtgcag cgatgggggc gggggggggg
     1561 ggggggcgcg cgccaggcgg ggcggggcgg ggcgaggggc ggggcggggc gaggcggaga
     1621 ggtgcggcgg cagccaatca gagcggcgcg ctccgaaagt ttccttttat ggcgaggcgg
     1681 cggcggcggc ggccctataa aaagcgaagc gcgcggcggg cgggagtcgc tgcgacgctg
     1741 ccttcgcccc gtgccccgct ccgccgccgc ctcgcgccgc ccgccccggc tctgactgac
     1801 cgcgttactc ccacaggtga gcgggcggga cggcccttct cctccgggct gtaattagct
     1861 gagcaagagg taagggttta agggatggtt ggttggtggg gtattaatgt ttaattacct
     1921 ggagcacctg cctgaaatca ctttttttca ggttggctcg aggccaccat gaaacggaca
     1981 gccgacggaa gcgagttcga gtcaccaaag aagaagcgga aagtctctga ggtggagttt
     2041 tcccacgagt actggatgag acatgccctg accctggcca agagggcacg ggatgagagg
     2101 gaggtgcctg tgggagccgt gctggtgctg aacaatagag tgatcggcga gggctggaac
     2161 agagccatcg gcctacacga cccaacagcc catgccgaaa ttatggccct gagacaaggc
     2221 ggcctggtca tgcagaacta cagactgatt gacgccaccc tgtacgtgac attcgagcct
     2281 tgcgtgatgt gcgctggcgc catgatccac tctaggatcg gccgcgtggt gtttggatgg
     2341 agaaattcta aaagaggcgc cgcaggctcc ctgatgaacg tgctgaacta ccccggcatg
     2401 aatcaccgcg tcgaaattac cgagggaatc ctggcagatg aatgtgccgc cctgctgtgc
     2461 gacttctatc ggatgcctag acaggtgttc aatgctcaga agaaggccca gagttccatc
     2521 aactccggag gatctagcgg aggctcctct ggcagcgaga caccaggaac aagcgagtca
     2581 gcaacaccag agagcagtgg cggcagcagc ggcggcagca tgatggccgt ggtgtacgtg
     2641 atcagcaagt ctggcaaacc cctgatgccc accacccggt gcggccacgt gagaatcctg
     2701 ctgaaggagg gcaaggccag agtggtggag cggaaaccgt tcaccatcca gctgacctac
     2761 gagagcgcgg aggaaacaca gcctctggtc ctgggcattg ctcctggacg caccaatatc
     2821 ggcatgagcg tggtgacaga atccggagag agcgtgttca acgcccagat cagaaccaga
     2881 aacaaggacg tgcctaagct gatgaaagac agaaagcagt accggatggc tcacagacgg
     2941 ctgaagcggc gctgcaagcg aagaagaaga gccaaggccg ccggcaccgc attcgaggaa
     3001 ggagaaaagc agagactgct gcctggctgt ttcaagccta tcacatgtaa gagcatcaga
     3061 aataaggaag ctagattcaa taacaggaag cggcctgtgg gatggctgac cccaaccgcc
     3121 aaccacctgc tggtgaccca cctgaacgtg gtcaagaagg tgcaaaagat cctgcctgtg
     3181 gccaaggtgg tgctggaact gaatcggttc agcttcatgg ccatgaacaa ccccaaggtg
     3241 cagcggtggc agtaccagag aggccctctg tacggtaaag gcagcgtgga agaggccgtg
     3301 agcatgcagc aggacggcca ctgcctgttt tgcaagcacg gcattgacca ctatcaccat
     3361 gtggttccta ggagaaagaa cggctctgaa accctggaga accgggtggg cctgtgcgag
     3421 gaacaccacc ggctcgtcca cacagataag gagtgggagg ccaatctggc ttctaagaag
     3481 agcggaatga acaagaaata ccacgccctg agcgtgctga accagatcat cccatacctg
     3541 gccgaccagc ttgccgatat gttccccggc aacttctgtg tgaccagcgg acaggataca
     3601 tacctgtttc gggaagaaca tggcatcccc aaggaccact acctggatgc ctactgcatc
     3661 gcctgcagcg ccctgaccga cgccaagaag gtgagcagcc ctaaaggcag accttacatg
     3721 gtgagacagt tcagacgcca cgatagacag gcctgtcaca aggccaacct gaacagaaga
     3781 tactacatgg gcggcaaact ggttgctaca aacagacata aggccatgga ccagaaaacc
     3841 gacagcctgg aggagtacag agccgcccac agcgccgctg acgtatccaa gctgacagtg
     3901 aagcaccctt ctgctcaata taaggacatg tcccggatca tgcctggcag catcctcgtg
     3961 tccggcgagg gcaagctctt tacactgaga agatccgagg gcagaaacaa gggccaagtg
     4021 aactacttcg tgagcaccga gggcatcaag tactgggcca gaaagtgcca gtatctgaga
     4081 aacaatggcg gcctgcagat ctacgtgtct ggaggatcta gcggaggatc caagagaacc
     4141 gctgatggct cagagtttga gagcccaaaa aagaaaagaa aagtcagtgg cggcagcagc
     4201 ggcggcagct ctgaggtgga gttttcccac gagtactgga tgagacatgc cctgaccctg
     4261 gccaagaggg cacgggatga gagggaggtg cctgtgggag ccgtgctggt gctgaacaat
     4321 agagtgatcg gcgagggctg gaacagagcc atcggcctac acgacccaac agcccatgcc
     4381 gaaattatgg ccctgagaca aggcggcctg gtcatgcaga actacagact gattgacgcc
     4441 accctgtacg tgacattcga gccttgcgtg atgtgcgctg gcgccatgat ccactctagg
     4501 atcggccgcg tggtgtttgg atggagaaat tctaaaagag gcgccgcagg ctccctgatg
     4561 aacgtgctga actaccccgg catgaatcac cgcgtcgaaa ttaccgaggg aatcctggca
     4621 gatgaatgtg ccgccctgct gtgcgacttc tatcggatgc ctagacaggt gttcaatgct
     4681 cagaagaagg cccagagttc catcaacaag agaaccgccg acggctccga gttcgagagc
     4741 cccaagaaga agcgcaaagt gtgagaattc ctagagctcg ctgatcagcc tcgactgtgc
     4801 cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag
     4861 gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta
     4921 ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag
     4981 agaatagcag gcatgctggg gagcggccgc tagttattaa tagtaatcaa ttacggggtc
     5041 attagttcat agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc
     5101 tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt
     5161 aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca
     5221 cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg
     5281 taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca
     5341 gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa
     5401 tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa
     5461 tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc
     5521 cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctgg
     5581 tttagtgaac cgtcagatcc gctagcgcca ccatggtgag caagggcgag gaggataaca
     5641 tggccatcat caaggagttc atgcgcttca aggtgcacat ggagggctcc gtgaacggcc
     5701 acgagttcga gatcgagggc gagggcgagg gccgccccta cgagggcacc cagaccgcca
     5761 agctgaaggt gaccaagggt ggccccctgc ccttcgcctg ggacatcctg tcccctcagt
     5821 tcatgtacgg ctccaaggcc tacgtgaagc accccgccga catccccgac tacttgaagc
     5881 tgtccttccc cgagggcttc aagtgggagc gcgtgatgaa cttcgatgac ggcggcgtgg
     5941 tgaccgtgac ccaggactcc tccctgcagg acggcgagtt catctacaag gtgaagctgc
     6001 gcggcaccaa cttcccctcc gacggccccg taatgcagaa gaaaaccatg ggctgggagg
     6061 cctcctccga gcggatgtac cccgaggacg gcgccctgaa gggcgagatc aagcagaggc
     6121 tgaagctgaa ggacggcggc cactacgacg ctgaggtcaa gaccacctac aaggccaaga
     6181 agcccgtgca gctgcccggc gcctacaacg tcaacatcaa gttggacatc acctcccaca
     6241 acgaggacta caccatcgtg gaacagtacg aacgcgccga gggccgccac tccaccggcg
     6301 gcatggacga gctgtataag tgatgtacac cgggatgcag aaattgatga tctattaaac
     6361 aataaagatg tccactaaaa tggaagtttt tcctgtcata ctttgttaag aagggtgaga
     6421 acagagtacc tacattttga atggaaggat tggagctacg ggggtggggg tggggtggga
     6481 ttagataaat gcctgctctt tactgaaggc tctttactat tgctttatga taatgtttca
     6541 tagttggata tcataattta aacaagcaaa accaaattaa gggccagctc attcctccca
     6601 ctcatgatct atagatctat agatctctcg tgggatcatt gtttttctct tgattcccac
     6661 tttgtggttc taagtactgt ggtttccaaa tgtgtcagtt tcatagcctg aagaacgaga
     6721 tcagcagcct ctgttccaca tacacttcat tctcagtatt gttttgccaa gttctaattc
     6781 catcagaagc tggtcgacct gcaggggcgc ctgatgcggt attttctcct tacgcatctg
     6841 tgcggtattt cacaccgcat acgtcaaagc aaccatagta cgcgccctgt agcggcgcat
     6901 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
     6961 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     7021 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     7081 ccaaaaaact tgatttgggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     7141 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     7201 caacactcaa ccctatctcg ggctattctt ttgatttata agggattttg ccgatttcgg
     7261 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat
     7321 taacgtttac aattttatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa
     7381 gccagccccg acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg
     7441 catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac
     7501 cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt ttataggtta
     7561 atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg
     7621 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     7681 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
     7741 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     7801 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
     7861 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
     7921 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag
     7981 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
     8041 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
     8101 tgagtgataa cactgcggcc aacttacttc tgacaacgat tggaggaccg aaggagctaa
     8161 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     8221 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa
     8281 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
     8341 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     8401 ggtttattgc tgataaatct ggagccggtg agcgtggaag ccgcggtatc attgcagcac
     8461 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     8521 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     8581 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
     8641 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     8701 agttttcgtt ccactgagcg tca
//
