LOCUS       Exported                6933 bp ds-DNA     circular SYN 17-SEP-2018
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 6933)
  AUTHORS   tanhua4444
  TITLE     Direct Submission
  JOURNAL   Exported Thursday, May 13, 2021 from SnapGene Viewer 5.2.5.1
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6933
                     /organism="recombinant plasmid"
                     /mol_type="other DNA"
     misc_feature    1..129
                     /label=ITR
     misc_feature    144..1149
                     /label=Insert
     promoter        144..1121
                     /label=CMV IE94 promoter
     primer_bind     1063..1080
                     /label=CMV-F
     promoter        1156..1174
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     CDS             1193..1255
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTGD"
     CDS             1256..1285
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1289..1435
                     /codon_start=1
                     /product="transmembrane domain from platelet derived growth
                     factor receptor beta"
                     /label=PDGFR-beta TM domain
                     /translation="AVGQDTQEVIVVPHSLPFKVVVISAILALVVLTIISLIILIMLWQ
                     KKPR"
     CDS             1436..1630
                     /codon_start=1
                     /label=Kv2.1
                     /translation="QSQPILNTKEMAPQSKPPEELEMSSMPSPVAPLPARTEGVIDMRS
                     MSSIDSFISCATDFPEATRF"
     CDS             1637..2083
                     /codon_start=1
                     /label=CaM
                     /translation="MPDQLTEEQIAEFKEAFSLFDKDGDGTITTKELGTVMRSLGQNPT
                     EAELQDMINEVDADGDGTIDFPEFLTMMARKMKYRDTEEEIREAFGVFDKDGNGYISAA
                     ELRHVMTNLGEKLTDEEVDEMIREADIDGDGQVNYEEFVQMMTAK"
     CDS             2156..2182
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     CDS             2186..2539
                     /codon_start=1
                     /label=TevN
                     /translation="MGESLFKGPRDYNPISSTICHLTNESDGHTTSLYGIGFGPFIITN
                     KHLFRRNNGTLLVQSLHGVFKVKNTTTLQQHLIDGRDMIIIRMPKDFPPFPQKLKFREP
                     QREERICLVTTNFQ"
     primer_bind     2500..2517
                     /label=TevN FW
     CDS             2561..2974
                     /codon_start=1
                     /label=AsLOV2
                     /translation="EFLATTLERIEKNFVITDPRLPDNPIIFASDSFLQLTEYSREEIL
                     GRNCRFLQGPETDRATVRKIRDAIDNQTEVTVQLINYTKSGKKFWNVFHLQPMRDYKGD
                     VQYFIGVQLDGTERLHGAAEREAVCLIKKTAFQI"
     CDS             2975..2995
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             2996..4000
                     /codon_start=1
                     /product="tetracycline-controlled transactivator, 
                     comprising a fusion of the tetracycline repressor TetR with
                     the C-terminal activation domain of herpes simplex virus 
                     VP16"
                     /label=tTA
                     /note="In the Tet-Off(R) system, tTA binds to the 
                     Tet-responsive element and stimulates transcription only in
                     the absence of tetracycline or doxycycline."
                     /translation="SRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHVK
                     NKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTRP
                     TEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTTD
                     SMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGSAYSRARTKNNYGSTIE
                     GLLDLPDDDAPEEAGLAAPRLSFLPAGHTRRLSTAPPTDVSLGDELHLDGEDVAMAHAD
                     ALDDFDLDMLGDGDSPGPGFTPHDSAPYGALDMADFEFEQMFTDALGIDEYGG"
     promoter        complement(4010..4028)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    4033..4167
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     repeat_region   4196..4336
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     rep_origin      4411..4866
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        5148..5252
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5253..6113
                     /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      6284..6872
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg ccctccagtg agcgagcgcg ccagagaggg agtggccaac tccatcacta
      121 ggggttcctg cggccgcacg cgtgccaatt caatatggcg tatatggact catgccaatt
      181 caatatggtg gatctggacc tgtgccaatt caatatggcg tatatggact cgtgccaatt
      241 caatatggtg gatctggacc ccagccaatt caatatggcg gacttggcac catgccaatt
      301 caatatggcg gacctggcac tgtgccaact ggggaggggt ctacttggca cggtgccaag
      361 tttgaggagg ggtcttggcc ctgtgccaag tccgccatat tgaattggca tggtgccaat
      421 aatggcggcc atattggcta tatgccagga tcaatatata ggcaatatcc aatatggccc
      481 tatgccaata tggctattgg ccaggttcaa tactatgtat tggccctatg ccatatagta
      541 ttccatatat gggttttcct attgacgtag atagcccctc ccaatgggcg gtcccatata
      601 ccatatatgg ggcttcctaa taccgcccat agccactccc ccattgacgt caatggtctc
      661 tatatatggt ctttcctatt gacgtcatat gggcggtcct attgacgtat atggcgcctc
      721 ccccattgac gtcaattacg gtaaatggcc cgcctggctc aatgcccatt gacgtcaata
      781 ggaccaccca ccattgacgt caatgggatg gctcattgcc cattcatatc cgttctcacg
      841 ccccctattg acgtcaatga cggtaaatgg cccacttggc agtacatcaa tatctattaa
      901 tagtaacttg gcaagtacat tactattgga agtacgccag ggtacattgg cagtactccc
      961 attgacgtca atggcggtaa atggcccgcg atggctgcca agtacatccc cattgacgtc
     1021 aatggggagg ggcaatgacg caaatgggcg ttccattgac gtaaatgggc ggtaggcgtg
     1081 cctaatggga ggtctatata agcaatgctc gtttagggaa ccgccattct gcctggggac
     1141 gtcggagcaa gcttgattta ggtgacacta tagaatacaa gctacttgtt ctatggagac
     1201 agacacactc ctgctatggg tactgctgct ctgggttcca ggttccactg gtgacgaaca
     1261 aaaactcatc tcagaagagg atctgaatgc tgtgggccag gacacgcagg aggtcatcgt
     1321 ggtgccacac tccttgccct ttaaggtggt ggtgatctca gccatcctgg ccctggtggt
     1381 gctcaccatc atctccctta tcatcctcat catgctttgg cagaagaagc cacgtcagtc
     1441 ccagcctatt ctgaacacca aagagatggc cccccagagt aaaccccctg aggagctgga
     1501 aatgtcctca atgccctccc ctgtggcacc tcttcctgca agaacagagg gcgtgatcga
     1561 catgaggtcc atgagttcta ttgactcttt tatctcatgc gccaccgatt ttccagaggc
     1621 tacgcggttt ctcgagatgc cggaccaact gactgaagag cagatcgcag aatttaaaga
     1681 ggctttctcc ctatttgaca aggacgggga tgggacaata acaaccaagg agctggggac
     1741 ggtgatgcgg tctctggggc agaaccccac agaagcagag ctgcaagaca tgatcaatga
     1801 agtagatgcc gacggtgacg gcacaatcga cttccctgag ttcctgacaa tgatggcaag
     1861 aaaaatgaaa tacagggaca cggaagaaga aattagagaa gcgttcggtg tgtttgataa
     1921 ggatggcaat ggctacatca gtgcagcaga gcttcgccac gtgatgacaa accttggaga
     1981 gaagttaaca gatgaagagg ttgatgaaat gatcagggaa gcagacatcg atggggatgg
     2041 tcaggtaaac tacgaagagt ttgtacaaat gatgacagcg aaggcatgcg gtggcggtgg
     2101 ctctggaggt ggtgggtccg gaggaggcgg ccgctccggt ggatccatgc ttcaacttcc
     2161 tcctcttgaa cgtcttactc tcgaaatggg cgagagcctg ttcaagggac ctagggacta
     2221 caaccctatc agtagcacaa tttgtcacct gaccaacgag agtgatggcc acacaacaag
     2281 cctgtacggc atcgggttcg gaccctttat catcaccaac aagcacctgt tcaggcggaa
     2341 taatggcact ctgctggtgc agagcctgca cggggtgttc aaagtgaaga acacaaccac
     2401 tctgcagcag cacctgatcg atgggcggga tatgatcatc attaggatgc ccaaggactt
     2461 cccccctttt cctcagaaac tgaagttccg agagccccag agagaggaga gaatctgtct
     2521 ggtgaccaca aactttcagg agctcggtag tggtagtggg gagtttctgg caaccacact
     2581 ggaacggatc gagaaaaatt tcgtgattac tgatccgaga ctgcctgaca acccaatcat
     2641 ttttgcgagc gattccttcc tgcagctgac agaatattct cgggaagaga tcctggggcg
     2701 caattgccgt tttctgcagg gacccgagac agaccgtgcc actgttcgga aaatcagaga
     2761 tgctattgac aaccagactg aagtgaccgt tcagctgatc aattatacca agagcggcaa
     2821 gaagttctgg aacgtgttcc acctgcagcc gatgcgcgat tataagggcg acgtccagta
     2881 cttcattggc gtgcagctgg atggcaccga acgtcttcat ggcgccgctg agcgtgaggc
     2941 ggtctgcctg atcaaaaaga cagcctttca gattgagaac ctgtacttcc agggctctag
     3001 attagataaa agtaaagtga ttaacagcgc attagagctg cttaatgagg tcggaatcga
     3061 aggtttaaca acccgtaaac tcgcccagaa gctaggtgta gagcagccta cattgtattg
     3121 gcatgtaaaa aataagcggg ctttgctcga cgccttagcc attgagatgt tagataggca
     3181 ccatactcac ttttgccctt tagaagggga aagctggcaa gattttttac gtaataacgc
     3241 taaaagtttt agatgtgctt tactaagtca tcgcgatgga gcaaaagtac atttaggtac
     3301 acggcctaca gaaaaacagt atgaaactct cgaaaatcaa ttagcctttt tatgccaaca
     3361 aggtttttca ctagagaatg cattatatgc actcagcgct gtggggcatt ttactttagg
     3421 ttgcgtattg gaagatcaag agcatcaagt cgctaaagaa gaaagggaaa cacctactac
     3481 tgatagtatg ccgccattat tacgacaagc tatcgaatta tttgatcacc aaggtgcaga
     3541 gccagccttc ttattcggcc ttgaattgat catatgcgga ttagaaaaac aacttaaatg
     3601 tgaaagtggg tccgcgtaca gccgcgcgcg tacgaaaaac aattacgggt ctaccatcga
     3661 gggcctgctc gatctcccgg acgacgacgc ccccgaagag gcggggctgg cggctccgcg
     3721 cctgtccttt ctccccgcgg gacacacgcg cagactgtcg acggcccccc cgaccgatgt
     3781 cagcctgggg gacgagctcc acttagacgg cgaggacgtg gcgatggcgc atgccgacgc
     3841 gctagacgat ttcgatctgg acatgttggg ggacggggat tccccgggtc cgggatttac
     3901 cccccacgac tccgccccct acggcgctct ggatatggcc gacttcgagt ttgagcagat
     3961 gtttaccgat gcccttggaa ttgacgagta cggtgggtag gctagcctcc ctatagtgag
     4021 tcgtattacg tagatccaga catgataaga tacattgatg agtttggaca aaccacaact
     4081 agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta
     4141 accattataa gctgcaataa acaagttggt aaccacgtgc ggaccgagcg gccgcaggaa
     4201 cccctagtga tggagttggc cactccctct ctgcgcgctc gctcgctcac tgaggccggg
     4261 cgaccaaagg tcgcccgacg cccgggcttt gcccgggcgg cctcagtgag cgagcgagcg
     4321 cgcagctgcc tgcaggggcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt
     4381 tcacaccgca tacgtcaaag caaccatagt acgcgccctg tagcggcgca ttaagcgcgg
     4441 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc
     4501 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
     4561 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
     4621 ttgatttggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
     4681 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
     4741 accctatctc gggctattct tttgatttat aagggatttt gccgatttcg gcctattggt
     4801 taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgttta
     4861 caattttatg gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagcccc
     4921 gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt
     4981 acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac
     5041 cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga
     5101 taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     5161 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     5221 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
     5281 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     5341 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
     5401 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
     5461 ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg
     5521 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
     5581 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     5641 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
     5701 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     5761 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca
     5821 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
     5881 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     5941 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     6001 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     6061 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
     6121 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     6181 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     6241 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     6301 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     6361 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     6421 caaatactgt ccttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     6481 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     6541 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     6601 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     6661 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     6721 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     6781 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     6841 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     6901 tcctggcctt ttgctggcct tttgctcaca tgt
//
