LOCUS       62056_21820        8122 bp DNA     circular SYN 10-SEP-2019
DEFINITION  Cloning vector pUbC-mNeonGreen4-tDeg, complete sequence.
ACCESSION   MN052907
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8122)
  AUTHORS   Wu J, Zaccara S, Khuperkar D, Kim H, Tanenbaum ME, Jaffrey SR.
  TITLE     Live imaging of mRNA using RNA-stabilized fluorogenic proteins
  JOURNAL   Nat. Methods 16 (9), 862-865 (2019)
  PUBMED    31471614
REFERENCE   2  (bases 1 to 8122)
  AUTHORS   Wu J, Zaccara S, Khuperkar D, Kim H, Tanenbaum ME, Jaffrey SR.
  TITLE     Direct Submission
  JOURNAL   Submitted (11-JUN-2019) Pharmacology, Weill Cornell Medicine, 1300 
            York Avenue, New York, NY 10022, United States
REFERENCE   3  (bases 1 to 8122)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat. 
            Methods"; date: "2019"; volume: "16"; issue: "9"; pages: "862-865"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (11-JUN-2019) Pharmacology, Weill Cornell Medicine, 1300 York 
            Avenue, New York, NY 10022, United States"
FEATURES             Location/Qualifiers
     source          1..8122
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        235..633
                     /label=UbC promoter
                     /note="human ubiquitin C promoter"
     CDS             646..1353
                     /codon_start=1
                     /label=mNeonGreen
                     /note="bright monomeric yellow-green fluorescent protein
                     derived from LanYFP (Shaner et al., 2013)"
                     /translation="MVSKGEEDNMASLPATHELHIFGSINGVDFDMVGQGTGNPNDGYE
                     ELNLKSTKGDLQFSPWILVPHIGYGFHQYLPYPDGMSPFQAAMVDGSGYQVHRTMQFED
                     GASLTVNYRYTYEGSHIKGEAQVKGTGFPADGPVMTNSLTAADWCRSKKTYPNDKTIIS
                     TFKWSYTTGNGKRYRSTARTTYTFAKPMAANYLKNQPMYVFRKTELKHSKTELNFKEWQ
                     KAFTDVMGMDELYK"
     polyA_signal    3719..3946
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3992..4420
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        4434..4763
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             4830..5621
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    5798..5931
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5968..5984)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(5992..6008)
                     /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(6016..6046)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(6061..6082)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(6370..6955)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(7129..7986)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(7987..8091)
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttggcctc
      241 cgcgccgggt tttggcgcct cccgcgggcg cccccctcct cacggcgagc gctgccacgt
      301 cagacgaagg gcgcagcgag cgtcctgatc cttccgcccg gacgctcagg acagcggccc
      361 gctgctcata agactcggcc ttagaacccc agtatcagca gaaggacatt ttaggacggg
      421 acttgggtga ctctagggca ctggttttct ttccagagag cggaacaggc gaggaaaagt
      481 agtcccttct cggcgattct gcggagggat ctccgtgggg cggtgaacgc cgatgattat
      541 ataaggacgc gccgggtgtg gcacagctag ttccgtcgca gccgggattt gggtcgcggt
      601 tcttgtttgt ggatcgctgt gatcgtcact tggaagcttg ccaccatggt gagcaagggc
      661 gaggaggata acatggcctc tctcccagcg acacatgagt tacacatctt tggctccatc
      721 aacggtgtgg actttgacat ggtgggtcag ggcaccggca atccaaatga tggttatgag
      781 gagttaaacc tgaagtccac caagggtgac ctccagttct ccccctggat tctggtccct
      841 catatcgggt atggcttcca tcagtacctg ccctaccctg acgggatgtc gcctttccag
      901 gccgccatgg tagatggctc cggataccaa gtccatcgca caatgcagtt tgaagatggt
      961 gcctccctta ctgttaacta ccgctacacc tacgagggaa gccacatcaa aggagaggcc
     1021 caggtgaagg ggactggttt ccctgctgac ggtcctgtga tgaccaactc gctgaccgct
     1081 gcggactggt gcaggtcgaa gaagacttac cccaacgaca aaaccatcat cagtaccttt
     1141 aagtggagtt acaccactgg aaatggcaag cgctaccgga gcactgcgcg gaccacctac
     1201 acctttgcca agccaatggc ggctaactat ctgaagaacc agccgatgta cgtgttccgt
     1261 aagacggagc tcaagcactc caagaccgag ctcaacttca aggagtggca aaaggccttt
     1321 accgatgtga tgggcatgga cgagctgtac aagggtggac atatgggcac agggtccaca
     1381 ggcggtaccg gcggagtttc caaaggagaa gaagacaata tggcatcact ccccgcaacc
     1441 cacgagttgc atattttcgg ttcaattaat ggagtagatt tcgatatggt tggccaggga
     1501 acaggaaacc caaacgacgg atatgaagag cttaatctca aaagtaccaa aggcgatctg
     1561 caattttctc cgtggatact cgtgccacac attggatacg gatttcacca atatctcccg
     1621 tatccggatg gaatgtcccc ctttcaagca gcaatggtgg acgggagtgg ttatcaggta
     1681 cacagaacca tgcagttcga ggacggggct tctctgaccg taaattatag gtatacttat
     1741 gaaggctcac atattaaggg cgaagcacag gttaaaggaa ccgggtttcc tgcggatggc
     1801 cccgtcatga ctaattctct gacagccgca gattggtgtc gctccaaaaa gacatacccg
     1861 aatgataaga ctataatctc aacattcaaa tggtcctata cgacaggcaa cgggaaacga
     1921 tatagatcca cggctcgaac aacttacaca ttcgctaaac ctatggccgc caattacctc
     1981 aaaaatcagc ccatgtatgt gtttaggaaa accgaattga agcattctaa aacggaactt
     2041 aattttaagg aatggcagaa ggctttcaca gacgtaatgg ggatggatga actctataaa
     2101 tcaggtctcg agtcctcagg gggaacgggt gggtccggag gagttagtaa aggtgaagag
     2161 gacaatatgg caagtttgcc tgcgactcac gagcttcata tctttgggtc tataaatggc
     2221 gttgacttcg atatggttgg ccaaggtact ggcaacccca atgacggtta cgaggagttg
     2281 aatctcaagt ccacaaaagg tgatcttcag ttcagccctt ggattctcgt acctcatatt
     2341 ggatatggct ttcaccagta ccttccatac ccagacggta tgtcaccctt tcaagctgcg
     2401 atggtggatg gttccggcta tcaggtccac cgaacgatgc aattcgagga cggggccagc
     2461 ctcaccgtta attataggta cacctatgag ggaagtcaca taaagggaga agcccaagtg
     2521 aaaggaacag gattcccagc tgatggtcca gtaatgacga actccttgac agcggctgac
     2581 tggtgtagaa gcaaaaagac gtatcctaat gacaagacca tcattagcac tttcaaatgg
     2641 agttatacca caggaaacgg caaacggtac agaagcactg ctagaactac ctacactttc
     2701 gcaaagccga tggctgcaaa ctatttgaag aatcagccca tgtacgtgtt tcgaaaaacg
     2761 gaacttaagc acagtaagac tgaacttaat ttcaaggagt ggcagaaggc gttcacggat
     2821 gtcatgggta tggatgaact gtataaggga gggtctggca ctgggggcac tgccagcagc
     2881 ggatccggtg gcggtgtgag caagggcgag gaggataaca tggcctctct cccagcgaca
     2941 catgagttac acatctttgg ctccatcaac ggtgtggact ttgacatggt gggtcagggc
     3001 accggcaatc caaatgatgg ttatgaggag ttaaacctga agtccaccaa gggtgacctc
     3061 cagttctccc cctggattct ggtccctcat atcgggtatg gcttccatca gtacctgccc
     3121 taccctgacg ggatgtcgcc tttccaggcc gccatggtag atggctccgg ataccaagtc
     3181 catcgcacaa tgcagtttga agatggtgcc tcccttactg ttaactaccg ctacacctac
     3241 gagggaagcc acatcaaagg agaggcccag gtgaagggga ctggtttccc tgctgacggt
     3301 cctgtgatga ccaactcgct gaccgctgcg gactggtgca ggtcgaagaa gacttacccc
     3361 aacgacaaaa ccatcatcag tacctttaag tggagttaca ccactggaaa tggcaagcgc
     3421 taccggagca ctgcgcggac cacctacacc tttgccaagc caatggcggc taactatctg
     3481 aagaaccagc cgatgtacgt gttccgtaag acggagctca agcactccaa gaccgagctc
     3541 aacttcaagg agtggcaaaa ggcctttacc gatgtgatgg gcatggacga gctgtacaag
     3601 ggcggaagat ccggtggtgg ttctggtcct cgtccccgtg gtactcgtgg taaaggtcgc
     3661 cgtattcgtc gccgcggtta atctagaggg cccgtttaaa cccgctgatc agcctcgact
     3721 gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg
     3781 gaaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct
     3841 gagtaggtgt cattctattc tggggggtgg gggtgggggc aggacagcaa gggggaggat
     3901 tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa
     3961 agaaccagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc attaagcgcg
     4021 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
     4081 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     4141 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     4201 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     4261 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     4321 aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg
     4381 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc
     4441 agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     4501 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     4561 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc
     4621 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     4681 atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt
     4741 ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt ttcggatctg
     4801 atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt
     4861 ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct
     4921 gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga
     4981 ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg
     5041 ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact
     5101 ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg
     5161 agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct
     5221 gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg
     5281 gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt
     5341 tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg
     5401 cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc
     5461 ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag
     5521 agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt
     5581 cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt
     5641 cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc
     5701 cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca
     5761 gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa
     5821 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     5881 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac
     5941 ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
     6001 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     6061 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     6121 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     6181 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     6241 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     6301 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     6361 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     6421 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     6481 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     6541 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     6601 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     6661 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     6721 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     6781 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     6841 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     6901 tagcggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     6961 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     7021 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     7081 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     7141 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     7201 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     7261 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     7321 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     7381 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     7441 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     7501 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     7561 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     7621 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     7681 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     7741 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     7801 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     7861 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     7921 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     7981 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     8041 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     8101 ccgaaaagtg ccacctgacg tc
//
