LOCUS       Exported                8612 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pUSE-Src-YF-UniRapR-mCerulean-myc
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8612)
  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 8612)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8612
                     /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(4202..4219)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4208..4432
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4478..4906
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4565..4584)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4775..4796
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4915..4935)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4920..5250
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5101..5236
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5163..5182
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5317..6111
                     /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(5371..5390)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5981..6000
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6285..6406
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6322..6341)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6376..6395
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6455..6471)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6455..6471)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6468..6490)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6479..6495
                     /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(6503..6533)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6548..6569
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6686..6703)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6857..7445)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6937..6956)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(7616..8476)
                     /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     8239..8258
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8477..8581)
                     /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 gcaccgggac 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 taccagcccg 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 tctaggcttc cgggtgttgc ctccgatgca cctgcctccc ttcatcctct ctctggggct
     4021 gcgttgcctc ggcaatgccc cttttggagc atggaagggc ttcagggatg gggcagcctg
     4081 gaaggacagg tggaggctgg tacagtgact gtcccatgcc tgccatgatc tgcagaattc
     4141 caccacactg gactagtgga tccgagctcg gtaccaagct taagtttaaa ccgctgatca
     4201 gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc
     4261 ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg
     4321 cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg
     4381 gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat ggcttctgag
     4441 gcggaaagaa ccagctgggg ctctaggggg tatccccacg cgccctgtag cggcgcatta
     4501 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
     4561 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     4621 gctctaaatc ggggcatccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     4681 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     4741 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     4801 acactcaacc ctatctcggt ctattctttt gatttataag ggattttggg gatttcggcc
     4861 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt ctgtggaatg
     4921 tgtgtcagtt agggtgtgga aagtccccag gctccccagg caggcagaag tatgcaaagc
     4981 atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga
     5041 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
     5101 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
     5161 tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga
     5221 ggcttttttg gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc
     5281 ggatctgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac
     5341 gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca
     5401 atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt
     5461 gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg
     5521 tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga
     5581 agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct
     5641 cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg
     5701 gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg
     5761 gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc
     5821 gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat
     5881 ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac
     5941 tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt
     6001 gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct
     6061 cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc
     6121 tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca
     6181 ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga
     6241 tcctccagcg cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag
     6301 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     6361 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac
     6421 cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt
     6481 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg
     6541 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt
     6601 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     6661 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     6721 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     6781 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     6841 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     6901 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     6961 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     7021 ttctcccttc gggaagcgtg gcgctttctc aatgctcacg ctgtaggtat ctcagttcgg
     7081 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     7141 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     7201 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     7261 tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc
     7321 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     7381 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     7441 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     7501 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     7561 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     7621 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     7681 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     7741 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     7801 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     7861 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     7921 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     7981 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     8041 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     8101 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     8161 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     8221 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     8281 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     8341 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     8401 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     8461 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     8521 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     8581 gcacatttcc ccgaaaagtg ccacctgacg tc
//
