LOCUS       62056_5705        7258 bp DNA     circular SYN 07-SEP-2020
DEFINITION  Cloning vector pcDNA3.1/AKAR, complete sequence.
ACCESSION   MT800777
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7258)
  AUTHORS   Zhang JZ, Lu TW, Stolerman LM, Tenner B, Yang JR, Zhang JF, Falcke 
            M, Rangamani P, Taylor SS, Mehta S, Zhang J.
  TITLE     Phase Separation of a PKA Regulatory Subunit Controls cAMP 
            Compartmentation and Oncogenic Signaling
  JOURNAL   Cell (2020) In press
  PUBMED    32846158
REFERENCE   2  (bases 1 to 7258)
  AUTHORS   Zhang JZ, Tenner B.
  TITLE     Direct Submission
  JOURNAL   Submitted (23-JUL-2020) Bioengineering, University of California San
            Diego, 9500 Gilman Dr, San Diego, CA 92093, USA
REFERENCE   3  (bases 1 to 7258)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Cell (2020)
            In press"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (23-JUL-2020) Bioengineering, University of California San Diego, 
            9500 Gilman Dr, San Diego, CA 92093, USA"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..7258
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        105..484
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        485..688
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     promoter        733..751
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             789..1499
                     /codon_start=1
                     /label=mRuby2
                     /note="monomeric red fluorescent protein derived from
                     mRuby, with improved photophysical properties (Lam et al., 
                     2012)"
                     /translation="MVSKGEELIKENMRMKVVMEGSVNGHQFKCTGEGEGNPYMGTQTM
                     RITVIEGGPLPFAFDILATSFMYGSRTFIKYPKGIPDFFKQSFPEGFTWERVTRYEDGG
                     VVTVMQDTSLEDGCLVYHVQVRGVNFPSNGPVMQKKTKGWEPNTEMMYPADGGLRGYTH
                     MALKVDGGGHLSCSFVTTYRSKKTVGNIKMPGIHAVDHRLERLEESDNEMFVVQREVAV
                     AKFAGLGGGMDELYK"
     CDS             2022..2663
                     /codon_start=1
                     /label=GFP(1-10)
                     /note="fragment of superfolder GFP consisting of the first
                     10 beta-strands"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATIGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG
                     KYKTRAVVKFEGDTLVNRIELKGTDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFTVRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQTVLSKDPNEK"
     polyA_signal    2725..2949
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2995..3423
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        3437..3766
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             3833..4624
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    4801..4934
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4971..4987)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(4995..5011)
                     /label=lac operator
                     /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(5019..5049)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(5064..5085)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(5373..5961)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6135..6992)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(6993..7097)
                     /label=AmpR promoter
ORIGIN
        1 ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc ttagggttag
       61 gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt gattattgac
      121 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
      181 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      241 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      301 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      361 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      421 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      481 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      541 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      601 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      661 acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca ctgcttactg
      721 gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc gtttaaactt
      781 aagcttggat ggtgtctaag ggcgaagagc tgatcaagga aaatatgcgt atgaaggtgg
      841 tcatggaagg ttcggtcaac ggccaccaat tcaaatgcac aggtgaagga gaaggcaatc
      901 cgtacatggg aactcaaacc atgaggatca cagtcatcga gggaggaccc ctgccatttg
      961 cctttgacat tcttgccacg tcgttcatgt atggcagccg tacttttatc aagtacccga
     1021 aaggcattcc tgatttcttt aaacagtcct ttcctgaggg ttttacttgg gaaagagtta
     1081 cgagatacga agatggtgga gtcgtcaccg tcatgcagga caccagcctt gaggatggct
     1141 gtctcgttta ccacgtccaa gtcagagggg taaactttcc ctccaatggt cccgtgatgc
     1201 agaagaagac caagggttgg gagcctaata cagagatgat gtatccagca gatggtggtc
     1261 tgaggggata cactcatatg gcactgaaag ttgatggtgg tggccatctg tcttgctctt
     1321 tcgtaacaac ttacaggtca aaaaagaccg tcgggaacat caagatgccc ggtatccatg
     1381 ccgttgatca ccgcctggaa aggttagagg aaagtgacaa tgaaatgttc gtagtacaac
     1441 gcgaagtcgc agttgccaag ttcgccgggc ttggtggtgg gatggacgag ctgtacaagc
     1501 gcatgcataa gttttctcaa gaacagatcg gcgaaaacat tgtgtgcagg gtcatttgta
     1561 ccacgggtca aattcccatc cgagatttgt cagctgatat ttcacaagtg cttaaggaaa
     1621 aacgatccat aaagaaagtt tggacatttg gtagaaaccc agcctgtgac tatcatttag
     1681 gaaacatttc aagactgtca aataagcatt tccaaatact actaggagaa gacggtaacc
     1741 ttttattgaa tgacatttcc actaatggga cctggttaaa tgggcaaaaa gtcgagaaga
     1801 acagcaatca gttactgtct caaggtgatg aaataaccgt tggtgtaggc gtggaatcag
     1861 atattttatc tctggtcatt ttcataaacg acaaatttaa gcagtgcctc gagcagaaca
     1921 aagttgatcg ctctgcaggt aagccaggca gcggcgaggg cagcaccaag ggcctgcgtc
     1981 gcgccaccct ggtggacggc ggcaccggcg gcagcgagct catgtccaaa ggagaagaac
     2041 tgtttaccgg tgttgtgcca attttggttg aactcgatgg tgatgtcaac ggacataagt
     2101 tctcagtgag aggcgaagga gaaggtgacg ccaccattgg aaaattgact cttaaattca
     2161 tctgtactac tggtaaactt cctgtaccat ggccgactct cgtaacaacg cttacgtacg
     2221 gagttcagtg cttttcgaga tacccagacc atatgaaaag acatgacttt tttaagtcgg
     2281 ctatgcctga aggttacgtg caagaaagaa caatttcgtt caaagatgat ggaaaatata
     2341 aaactagagc agttgttaaa tttgaaggag atactttggt taaccgcatt gaactgaaag
     2401 gaacagattt taaagaagat ggtaatattc ttggacacaa actcgaatac aattttaata
     2461 gtcataacgt atacatcact gctgataagc aaaagaacgg aattaaagcg aatttcacag
     2521 tacgccataa tgtagaagat ggcagtgttc aacttgccga ccattaccaa caaaacaccc
     2581 ctattggaga cggtccggta cttcttcctg ataatcacta cctctcaaca caaacagtcc
     2641 tgagcaaaga tccaaatgaa aaaggaacag gtggcggcgg aagttagtct agagggcccg
     2701 tttaaacccg ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc
     2761 cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa
     2821 atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg
     2881 ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg
     2941 gctctatggc ttctgaggcg gaaagaacca gctggggctc tagggggtat ccccacgcgc
     3001 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     3061 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     3121 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
     3181 tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc
     3241 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
     3301 tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga
     3361 ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
     3421 attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg
     3481 cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg
     3541 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc
     3601 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
     3661 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt
     3721 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc
     3781 ttatatatcc attttcggat ctgatcaaga gacaggatga ggatcgtttc gcatgattga
     3841 acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga
     3901 ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg
     3961 gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga
     4021 ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt
     4081 tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct
     4141 gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct
     4201 gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg
     4261 agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca
     4321 ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga
     4381 tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt
     4441 ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt
     4501 ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct
     4561 ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt
     4621 cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca
     4681 cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg
     4741 gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc
     4801 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
     4861 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
     4921 tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg
     4981 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
     5041 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
     5101 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     5161 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     5221 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     5281 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     5341 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     5401 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     5461 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     5521 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     5581 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     5641 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     5701 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     5761 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     5821 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     5881 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     5941 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     6001 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     6061 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     6121 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     6181 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     6241 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     6301 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     6361 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     6421 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     6481 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     6541 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     6601 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     6661 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     6721 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     6781 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     6841 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     6901 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     6961 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     7021 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     7081 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcga cggatcggga
     7141 gatctcccga tcccctatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa
     7201 gccagtatct gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaat
//
