LOCUS       Exported                8025 bp DNA     circular SYN 24-JAN-2022
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 8025)
  AUTHORS   Jin Zhang Lab
  TITLE     Direct Submission
  JOURNAL   Exported Mar 14, 2022 from SnapGene 6.0.2
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8025
                     /mol_type="other DNA"
                     /organism="recombinant plasmid"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
                     /note="color: #ffffff"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
                     /note="color: #ffffff; direction: RIGHT"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /note="color: #ffffff; direction: RIGHT"
     CDS             904..2115
                     /codon_start=1
                     /label=PKA RIIalpha
                     /note="color: #5e5e5e"
                     /translation="MSHIQIPPGLTELLQGYTVEVLRQQPPDLVEFAVEYFTRLREARA
                     PASVLPAATPRQSLGHPPPEPGPDRVADAKGDSESEEDEDLEVPVPSRFNRRVSVCAET
                     YNPDEEEEDTDPRVIHPKTDEQRCRLQEACKDILLFKNLDQEQLSQVLDAMFERIVKAD
                     EHVIDQGDDGDNFYVIERGTYDILVTKDNQTRSVGQYDNRGSFGELALMYNTPRAATIV
                     ATSEGSLWGLDRVTFRRIIVKNNAKKRKMFESFIESVPLLKSLEVSERMKIVDVIGEKI
                     YKDGERIITQGEKADSFYIIESGEVSILIRSRTKSNKDGGNQEVEIARCHKGQYFGELA
                     LVTNKPRAASAYAVGDVKCLVMDVQAFERLLGPCMDIMKRNISHYEEQLVKMFGSSVDL
                     GNLGQ"
     CDS             2125..2796
                     /codon_start=1
                     /label=Dronpa (H193T)
                     /note="color: #008f00"
                     /translation="MSVIKPDMKIKLRMEGAVNGHPFAIEGVGLGKPFEGKQSMDLKVK
                     EGGPLPFAYDILTTVFCYGNRVFAKYPENIVDYFKQSFPEGYSWERSMNYEDGGICNAT
                     NDITLDGDCYIYEIRFDGVNFPANGPVMQKRTVKWEPSTEKLYVRDGVLKGDVNMALSL
                     EGGGHYRCDFKTTYKAKKVVQLPDYHFVDTHIEIKSHDKDYSNVNLHEHAEAHSELPRQ
                     AK"
     CDS             2821..3513
                     /codon_start=1
                     /label=TagRFP-T
                     /note="color: #ff2600"
                     /translation="MSELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTMRIKV
                     VEGGPLPFAFDILATSFMYGSRTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEANTEMLYPADGGLEGRTDMALK
                     LVGGGHLICNFKTTYRSKKPAKNLKMPGVYYVDHRLERIKEADKETYVEQHEVAVARYC
                     DLPSKLGHK"
     promoter        complement(3577..3595)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
                     /note="color: #ffffff; direction: LEFT"
     polyA_signal    3621..3845
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
                     /note="color: #a6acb3"
     rep_origin      3891..4319
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
                     /note="color: #ffff00"
     promoter        4333..4663
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
                     /note="color: #ffffff; direction: RIGHT"
     rep_origin      4514..4649
                     /label=SV40 ori
                     /note="SV40 origin of replication"
                     /note="color: #ffff00"
     CDS             4730..5524
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /note="color: #ccffcc"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    5698..5819
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
                     /note="color: #a6acb3"
     primer_bind     complement(5868..5884)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
                     /note="color: #a020f0; direction: LEFT"
     protein_bind    5892..5908
                     /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)."
                     /note="color: #31849b"
     promoter        complement(5916..5946)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
                     /note="This reverse directional feature has 3 segments:
                        1: 5916 .. 5922 / #ffffff / -10
                        2: 5923 .. 5940 / #ffffff
                        3: 5941 .. 5946 / #ffffff / -35"
     protein_bind    5961..5982
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
                     /note="color: #31849b"
     rep_origin      complement(6270..6858)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #ffff00"
     CDS             complement(7029..7889)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /note="This feature has 2 segments:
                        1: 7029 .. 7820 / #ccffcc
                        2: 7821 .. 7889 / #ccffcc / signal sequence
                     Cleavage site after base 7820"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(7890..7994)
                     /gene="bla"
                     /label=AmpR promoter
                     /note="color: #ffffff; direction: LEFT"
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtcgactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggac tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gcttgccgcc
      901 accatgagcc acatccagat cccgccgggg ctcacggagc tgctgcaggg ctacacggtg
      961 gaggtgctgc gacagcagcc gcctgacctc gtcgaattcg cagtggagta cttcacccgc
     1021 ctgcgcgagg cccgcgcccc agcctcagtc ctgcccgccg ccaccccacg ccagagcctg
     1081 ggccaccccc cgccagaacc cggcccggac cgtgtcgccg acgccaaagg ggacagcgag
     1141 tcggaggagg acgaggactt ggaagttcca gttcctagca gatttaatag acgagtatca
     1201 gtctgtgctg agacctataa ccctgatgag gaagaggaag atacagatcc aagggtgatt
     1261 catcctaaaa ctgatgaaca gagatgcaga cttcaggaag cttgcaaaga tattctcctt
     1321 ttcaaaaatc ttgatcagga acagctttct caagttctcg atgccatgtt tgaaaggata
     1381 gtcaaagctg atgagcatgt cattgaccaa ggagatgatg gagacaactt ttatgtcata
     1441 gaacggggaa cttatgacat tttagtaaca aaagataatc aaacccgctc tgttggtcaa
     1501 tatgacaacc gtggcagttt tggagaacta gctctgatgt acaacacccc gagagctgct
     1561 accattgttg ctacctcaga aggctccctt tggggactgg accgggtgac ttttagaaga
     1621 atcatagtga aaaataatgc aaagaagagg aagatgtttg aatcatttat tgagtctgtg
     1681 cccctcctta aatcactaga ggtgtcagaa cgaatgaaga ttgtggatgt aataggagag
     1741 aagatctata aggatggaga acgcataatc actcagggtg aaaaggctga tagcttttac
     1801 atcatagagt ctggcgaagt gagcatcttg attagaagca ggactaaatc aaacaaggat
     1861 ggtgggaacc aggaggtcga gattgcccgc tgccataagg ggcagtactt tggagagctt
     1921 gccctggtca ccaacaaacc cagagctgcc tcagcttatg cagttggaga tgtcaaatgc
     1981 ttagttatgg atgtacaagc attcgagagg cttctggggc cctgcatgga catcatgaag
     2041 aggaacatct cacactatga ggaacagctg gtgaagatgt ttggctccag cgtggatctg
     2101 ggcaacctcg ggcaggggga tcccatgagt gtgattaaac cagacatgaa gatcaagctg
     2161 cgtatggaag gcgctgtaaa tggacacccg ttcgcgattg aaggagttgg ccttgggaag
     2221 cctttcgagg gaaaacagag tatggacctt aaagtcaaag aaggcggacc tctgcctttc
     2281 gcctatgaca tcttgacaac tgtgttctgt tacggcaaca gggtattcgc caaataccca
     2341 gaaaatatag tagactattt caagcagtcg tttcctgagg gctactcttg ggaacgaagc
     2401 atgaattacg aagacggggg catttgtaac gcgacaaacg acataaccct ggatggtgac
     2461 tgttatatct atgaaattcg atttgatggt gtgaactttc ctgccaatgg tccagttatg
     2521 cagaagagga ctgtgaaatg ggagccatcc actgagaaat tgtatgtgcg tgatggagtg
     2581 ctgaagggtg atgttaacat ggctctgtcg cttgaaggag gtggccatta ccgatgtgac
     2641 ttcaaaacta cttataaagc taagaaggtt gtccagttgc cagactatca ctttgtggac
     2701 acacacattg agattaaaag ccacgacaaa gattacagta atgttaatct gcatgagcat
     2761 gccgaagcgc attctgagct gccgaggcag gccaagagat ctgacgaggt ggacggtacc
     2821 atgagcgagc tgattaagga gaacatgcac atgaagctgt acatggaggg caccgtgaac
     2881 aaccaccact tcaagtgcac atccgagggc gaaggcaagc cctacgaggg cacccagacc
     2941 atgagaatca aggtggtcga gggcggccct ctccccttcg ccttcgacat cctggctacc
     3001 agcttcatgt acggcagcag aaccttcatc aaccacaccc agggcatccc cgatttcttt
     3061 aagcagtcct tccctgaggg cttcacatgg gagagagtca ccacatacga agacgggggc
     3121 gtgctgaccg ctacccagga caccagcctc caggacggct gcctcatcta caacgtcaag
     3181 atcagagggg tgaacttccc atccaacggc cctgtgatgc agaagaaaac actcggctgg
     3241 gaggccaaca ccgagatgct gtaccccgct gacggcggcc tggaaggcag aaccgacatg
     3301 gccctgaagc tcgtgggcgg gggccacctg atctgcaact tcaagaccac atacagatcc
     3361 aagaaacccg ctaagaacct caagatgccc ggcgtctact atgtggacca cagactggaa
     3421 agaatcaagg aggccgacaa agagacctac gtcgagcagc acgaggtggc tgtggccaga
     3481 tactgcgacc tccctagcaa actggggcac aagtgagaat tctgcagata tccatcacac
     3541 tggcggccgc tcgagcatgc atctagaggg ccctattcta tagtgtcacc taaatgctag
     3601 agctcgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc
     3661 ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga
     3721 ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca
     3781 ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc
     3841 tatggcttct gaggcggaaa gaaccagctg gggctctagg gggtatcccc acgcgccctg
     3901 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     3961 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     4021 ctttccccgt caagctctaa atcggggcat ccctttaggg ttccgattta gtgctttacg
     4081 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg
     4141 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     4201 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt
     4261 ggggatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta
     4321 attctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc aggcaggcag
     4381 aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc
     4441 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc
     4501 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
     4561 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg agctattcca
     4621 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc cgggagcttg
     4681 tatatccatt ttcggatctg atcaagagac aggatgagga tcgtttcgca tgattgaaca
     4741 agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg
     4801 ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg
     4861 cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aggacgaggc
     4921 agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt
     4981 cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc
     5041 atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca
     5101 tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc
     5161 acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg
     5221 gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc atgcccgacg gcgaggatct
     5281 cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc
     5341 tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc
     5401 tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta
     5461 cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt
     5521 ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga
     5581 gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac
     5641 gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccccaac
     5701 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
     5761 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat
     5821 catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc atagctgttt
     5881 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
     5941 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
     6001 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     6061 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     6121 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     6181 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     6241 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     6301 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     6361 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     6421 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcaatgctc acgctgtagg
     6481 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     6541 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     6601 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     6661 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt
     6721 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     6781 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     6841 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     6901 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     6961 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     7021 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     7081 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     7141 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     7201 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     7261 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     7321 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     7381 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     7441 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     7501 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     7561 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     7621 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     7681 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     7741 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     7801 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     7861 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     7921 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     7981 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtc
//
