LOCUS       Exported                3746 bp ds-DNA     circular SYN 04-DEC-2018
DEFINITION  MoClo golden gate assembly CD part for NahR-sfGFP fusion (Adam Meyer
            improved NahR (C75m) fused to superfolder GFP (C3m))..
ACCESSION   .
VERSION     .
KEYWORDS    C90m
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3746)
  TITLE     CIDAR MoClo Extension
REFERENCE   2  (bases 1 to 3746)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 4, 2018 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     misc_feature    3727..2080
                     /ApEinfo_revcolor="#6ecff6"
                     /ApEinfo_fwdcolor="#6ecff6"
                     /label="DVLA"
     source          <2082..>2981
                     /organism="recombinant plasmid"
                     /mol_type="other DNA"
                     /label="source recombinant plasmid"
     CDS             2082..3722
                     /created_by="murraylab"
                     /label="NahR sfGFP"
     misc_feature    2099
                     /label="T"
                     /note="Geneious type: Editing History Replacement"
     mutation        2154..2156
                     /label="I25V"
     mutation        2295..2297
                     /label="P72H"
     mutation        2433..2435
                     /label="V118A"
     misc_feature    2434
                     /label="T"
                     /note="Geneious type: Editing History Replacement"
     mutation        2823..2825
                     /label="R425C"
     linker          2982..3005
                     /created_by="User"
                     /modified_by="murraylab"
                     /label="GS linker"
     linker          2982..3005
                     /created_by="User"
                     /modified_by="murraylab"
                     /label="GS linker"
     source          1..3746
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     complement(14..31)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(185..773)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(265..284)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(944..1804)
                     /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     1567..1586
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1805..1909)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1977..1995
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      1999..2042
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    2045..2066
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     CDS             3006..3719
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     misc_feature    3734..8
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
ORIGIN
        1 cgctgcaggc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
       61 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
      121 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
      181 cgtttttcca caggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
      241 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
      301 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
      361 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
      421 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
      481 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
      541 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
      601 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
      661 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
      721 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
      781 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
      841 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
      901 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
      961 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     1021 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     1081 cgcgcgaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     1141 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     1201 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     1261 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     1321 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     1381 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     1441 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     1501 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     1561 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     1621 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     1681 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     1741 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     1801 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     1861 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     1921 gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata
     1981 ggcgtatcac gaggcagaat ttcagataaa aaaaatcctt agctttcgct aaggatgatt
     2041 tctggaattc gcggccgctt ctagagacta gtgggtctca aatggaactg cgtgacctgg
     2101 atttaaacct gctggtggtg ttcaaccagt tgctggtcga cagacgcgtc tctgtcactg
     2161 cggagaacct gggcctgacc cagcctgccg tgagcaatgc gctgaaacgc ctgcgcacct
     2221 cgctacagga cccactcttc gtgcgcacac atcagggaat ggaacccaca ccctatgccg
     2281 cgcatctggc cgagcacgtc acttcggcca tgcacgcact gcgcaacgcc ctacagcacc
     2341 atgaaagctt cgatccgctg accagcgagc gtaccttcac cctggccatg accgacattg
     2401 gcgagatcta cttcatgccg cggctgatgg atgcgctggc tcaccaggcc cccaattgcg
     2461 tgatcagtac ggtgcgcgac agttcgatga gcctgatgca ggccttgcag aacggaaccg
     2521 tggacttggc cgtgggcctg cttcccaatc tgcaaactgg cttctttcag cgccggctgc
     2581 tccagaatca ctacgtgtgc ctatgtcgca aggaccatcc agtcacccgc gaacccctga
     2641 ctctggagcg cttctgttcc tacggccacg tgcgtgtcat cgccgctggc accggccacg
     2701 gcgaggtgga cacgtacatg acacgggtcg gcatccggcg cgacatccgt ctggaagtgc
     2761 cgcacttcgc cgccgttggc cacatcctcc agcgcaccga tctgctcgcc actgtgccga
     2821 tatgtttagc cgactgctgc gtagagccct tcggcctaag cgccttgccg cacccagtcg
     2881 tcttgcctga aatagccatc aacatgttct ggcatgcgaa gtaccacaag gacctagcca
     2941 atatttggtt gcggcaactg atgtttgacc tgtttacgga tggaagcgga agtggatccg
     3001 gtagtatgcg taaaggcgaa gagctgttca ctggtgtcgt ccctattctg gtggaactgg
     3061 atggtgatgt caacggtcat aagttttccg tgcgtggcga gggtgaaggt gacgcaacta
     3121 atggtaaact gacgctgaag ttcatctgta ctactggtaa actgccggta ccttggccga
     3181 ctctggtaac gacgctgact tatggtgttc agtgctttgc tcgttatccg gaccatatga
     3241 agcagcatga cttcttcaag tccgccatgc cggaaggcta tgtgcaggaa cgcacgattt
     3301 cctttaagga tgacggcacg tacaaaacgc gtgcggaagt gaaatttgaa ggcgataccc
     3361 tggtaaaccg cattgagctg aaaggcattg actttaaaga agatggcaat atcctgggcc
     3421 ataagctgga atacaatttt aacagccaca atgtttacat caccgccgat aaacaaaaaa
     3481 atggcattaa agcgaatttt aaaattcgcc acaacgtgga ggatggcagc gtgcagctgg
     3541 ctgatcacta ccagcaaaac actccaatcg gtgatggtcc tgttctgctg ccagacaatc
     3601 actatctgag cacgcaaagc gttctgtcta aagatccgaa cgagaaacgc gatcatatgg
     3661 ttctgctgga gttcgtaacc gcagcgggca tcacgcatgg tatggatgaa ctgtacaaat
     3721 aaaggtagag acctactagt agcggc
//
