LOCUS       Exported                8674 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pUSE-Src-KD-UniRapR-mCerulean-myc
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8674)
  AUTHORS   Dagliyan O, Shirvanyants D, Karginov AV, Ding F, Fee L, 
            Chandrasekaran SN, Freisinger CM, Smolen GA, Huttenlocher A, Hahn 
            KM, Dokholyan NV
  TITLE     Rational design of a ligand-controlled protein conformational 
            switch.
  JOURNAL   Proc Natl Acad Sci U S A. 2013 Apr 23;110(17):6800-4. doi: 
            10.1073/pnas.1218319110. Epub 2013 Apr 8.
  PUBMED    23569285
REFERENCE   2  (bases 1 to 8674)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8674
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             3174..3890
                     /codon_start=1
                     /product="enhanced monomeric variant of CFP (Rizzo et al., 
                     2004)"
                     /label=mCerulean
                     /note=" mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNAISDNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3219..3240)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3480..3499)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3827..3848
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             3903..3932
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(4264..4281)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4270..4494
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4540..4968
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4627..4646)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4837..4858
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4977..4997)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4982..5312
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5163..5298
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5225..5244
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5379..6173
                     /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))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5433..5452)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6043..6062
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6347..6468
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6384..6403)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6438..6457
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6517..6533)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6517..6533)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6530..6552)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6541..6557
                     /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)."
     promoter        complement(6565..6595)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6610..6631
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6748..6765)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6919..7507)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6999..7018)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(7678..8538)
                     /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"
     primer_bind     8301..8320
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8539..8643)
                     /gene="bla"
                     /label=AmpR promoter
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 gctggctagc
      901 gtttaaacgg gccctctaga ctcgagcggc cgccactgtg ctggatcatg ggcagcaaca
      961 agagcaagcc caaggacgcc agccagcggc gccgcagcct ggagccctcg gaaaacgtgc
     1021 acggggcagg gggcgccttc ccggcctcac agacaccgag caagcccgcc tccgccgacg
     1081 gccaccgcgg gcccagcgcc gccttcgtgc cgcccgcggc cgagcccaag ctcttcggag
     1141 gcttcaactc ctcggacacc gtcacctccc cgcagagggc ggggcctctg gcaggtgggg
     1201 tgaccacctt tgtggccctc tatgactatg agtcacggac agagactgac ctgtccttca
     1261 agaaagggga gcggctgcag attgttaaca acacggaggg agactggtgg ctggcacact
     1321 cgctgagcac gggacagacc ggttacatcc ccagcaacta tgtggcgccc tccgactcca
     1381 tccaggctga ggagtggtac tttggcaaga tcactagacg ggaatcagag cggctgctgc
     1441 tcaacgccga gaacccgaga gggaccttcc tcgtgaggga gagtgagacc acaaaaggtg
     1501 cctactgcct ctctgtatcc gacttcgaca atgccaaggg cctaaatgtg aaacactaca
     1561 agatccgcaa gctggacagc ggcggtttct acatcacctc ccgcacccag ttcaacagcc
     1621 tgcagcagct cgtggcttac tactccaaac atgctgatgg cctgtgtcac cgcctcacta
     1681 ccgtatgtcc cacatccaag cctcagaccc agggattggc caaggatgcg tgggagatcc
     1741 cccgggagtc cctgcggctg gaggtcaagc tgggccaggg ttgcttcgga gaggtgtgga
     1801 tggggacctg gaacggacca ggtacctgcg tggtgcacta caccgggatg cttgaagatg
     1861 gaaagaaatt tgattcctcc cgggacagaa acaagccctt taagtttatg ctaggcaagc
     1921 aggaggtgat ccgaggctgg gaagaagggg ttgcccagat gagtgtgggt cagagagcca
     1981 aactgactat atctccagat tatgcctatg gtgccactgg gcacggttcg ggctccggat
     2041 caggcgtcaa ggacctcctc caagcctggg acctctatta tcatgtgttc cgacgaatct
     2101 caggtcctcc aggacctgga tcaggtctct ggcatgagat gtggcatgaa ggcctggaag
     2161 aggcatctcg tttgtacttt ggggaaagga acgtgaaagg catgtttgag gtgctggagc
     2221 ccttgcatgc tatgatggaa cggggccccc agactctgaa ggaaacatcc tttaatcagg
     2281 cctatggtcg agatttaatg gaggcccaag agtggtgcag gaagtacatg aaatcagggt
     2341 catcaggggg ctccggatca ggcatcatcc caccacatgc cactctcgtc ttcgatgtgg
     2401 agcttctaaa actggaaggt cccggaacca cgagggttgc catcaaaact ctgaagccag
     2461 gcaccatgtc cccagaggcc ttcctgcagg aggcccaagt catgaagaaa ctgaggcacg
     2521 agaaactggt gcagctgtat gctgtggtgt cggaagaacc catttacatt gtgacagagt
     2581 acatgaacaa ggggagtctg ctggactttc tcaaggggga aacgggcaaa tatttgcggc
     2641 taccccagct ggtggacatg tctgctcaga tcgcttcagg catggcctat gtggagcgga
     2701 tgaactatgt gcaccggcgc cttcgagccg ccaatatcct agtaggggag aacctggtgt
     2761 gcaaagtggc cgactttggg ttggcccggc tcatagaaga caacgaatac acagcccggc
     2821 aaggtgccaa attccccatc aagtggaccg cccctgaagc tgctctgtac ggcaggttca
     2881 ccatcaagtc ggatgtgtgg tcctttggga ttctgctgac cgagctcacc actaagggaa
     2941 gagtgcccta tcctgggatg gtgaaccgtg aggttctgga ccaggtggag cggggctacc
     3001 ggatgccttg tccccccgag tgccccgagt ccctgcatga ccttatgtgc cagtgctggc
     3061 ggaaggagcc cgaggagcgg cccaccttcg agtacctgca ggccttcctg gaagactact
     3121 ttacgtccac tgagccacag ttccagcccg gggagaacct aggcggacca ggaatggtga
     3181 gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg
     3241 taaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc tacggcaagc
     3301 tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc accctcgtga
     3361 ccaccctgac ctggggcgtg cagtgcttcg cccgctaccc cgaccacatg aagcagcacg
     3421 acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc ttcttcaagg
     3481 acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc
     3541 gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg
     3601 agtacaacgc catcagcgac aacgtctata tcaccgccga caagcagaag aacggcatca
     3661 aggccaactt caagatccgc cacaacatcg aggacggcag cgtgcagctc gccgaccact
     3721 accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac cactacctga
     3781 gcacccagtc caagctgagc aaagacccca acgagaagcg cgatcacatg gtcctgctgg
     3841 agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtacaag tccggaagcg
     3901 gggagcagaa gctgatctcc gaggaggacc tgtgaggact gtgtgcgcag aactgtcacc
     3961 tctagagcca cagtaccagc ccggggagaa cctataggga ctgtgtgcgc agaactgtca
     4021 cctctaggct tccgggtgtt gcctccgatg cacctgcctc ccttcatcct ctctctgggg
     4081 ctgcgttgcc tcggcaatgc cccttttgga gcatggaagg gcttcaggga tggggcagcc
     4141 tggaaggaca ggtggaggct ggtacagtga ctgtcccatg cctgccatga tctgcagaat
     4201 tccaccacac tggactagtg gatccgagct cggtaccaag cttaagttta aaccgctgat
     4261 cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt
     4321 ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat
     4381 cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg
     4441 gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcttctg
     4501 aggcggaaag aaccagctgg ggctctaggg ggtatcccca cgcgccctgt agcggcgcat
     4561 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
     4621 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     4681 aagctctaaa tcggggcatc cctttagggt tccgatttag tgctttacgg cacctcgacc
     4741 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     4801 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     4861 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg gggatttcgg
     4921 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa
     4981 tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca ggcaggcaga agtatgcaaa
     5041 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca
     5101 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
     5161 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     5221 tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag
     5281 gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt
     5341 tcggatctga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc
     5401 acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga
     5461 caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt
     5521 ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat
     5581 cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg
     5641 gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg
     5701 ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc
     5761 cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga
     5821 tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag
     5881 ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc
     5941 atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg
     6001 actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata
     6061 ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg
     6121 ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac
     6181 tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc
     6241 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat
     6301 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc
     6361 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     6421 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat
     6481 accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     6541 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     6601 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     6661 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     6721 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     6781 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     6841 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     6901 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     6961 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     7021 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     7081 ctttctccct tcgggaagcg tggcgctttc tcaatgctca cgctgtaggt atctcagttc
     7141 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     7201 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     7261 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     7321 gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc
     7381 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     7441 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     7501 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     7561 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     7621 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     7681 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     7741 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     7801 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     7861 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     7921 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     7981 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     8041 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     8101 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     8161 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     8221 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     8281 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     8341 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     8401 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     8461 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     8521 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     8581 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     8641 gcgcacattt ccccgaaaag tgccacctga cgtc
//
