LOCUS       Exported                5796 bp ds-DNA     circular SYN 23-NOV-2017
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 5796)
  AUTHORS   SILS-BCB
  TITLE     Direct Submission
  JOURNAL   Exported Sunday, Feb 24, 2019 from SnapGene 4.3.0
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5796
                     /organism="recombinant plasmid"
                     /mol_type="other DNA"
     misc_feature    complement(190..195)
                     /label=-35_Signal
                     /label=-35_Signal(trp)_1(2)
     misc_feature    1186..1215
                     /label=Promotor_P_1
     misc_feature    1186..1191
                     /label=-35_Signal
                     /label=-35_Signal(trp)_1
     misc_feature    1209..1215
                     /label=-10_Signal
                     /label=-10_Signal(lacUV5)_1
     misc_binding    1219..1241
                     /label=LacO
     misc_feature    1221..1241
                     /label=Protein_Binding_1
     misc_feature    1246..1252
                     /label=RBS
     primer_bind     1259..1277
                     /label=nm235
     CDS             1260..1967
                     /codon_start=1
                     /product="bright monomeric yellow-green fluorescent protein
                     derived from LanYFP (Shaner et al., 2013)"
                     /label=mNeonGreen
                     /translation="MVSKGEEDNMASLPATHELHIFGSINGVDFDMVGQGTGNPNDGYE
                     ELNLKSTKGDLQFSPWILVPHIGYGFHQYLPYPDGMSPFQAAMVDGSGYQVHRTMQFED
                     GASLTVNYRYTYEGSHIKGEAQVKGTGFPADGPVMTNSLTAADWCRSKKTYPNDKTIIS
                     TFKWSYTTGNGKRYRSTARTTYTFAKPMAANYLKNQPMYVFRKTELKHSKTELNFKEWQ
                     KAFTDVMGMDELYK"
     primer_bind     1260..1280
                     /label=XhoI-linker-mCherry
     primer_bind     1260..1277
                     /label=nm73
     primer_bind     1260..1276
                     /label=nm39
     variation       1263..1289
                     /label=Dispensible
     misc_feature    1296..1298
                     /label=L>CTG
     misc_feature    1299..1301
                     /label=P>CCG
     misc_feature    1305..1307
                     /label=T>ACC
     misc_feature    1314..1316
                     /label=L>CTG
     misc_feature    1377..1379
                     /label=P>CCG
     misc_feature    1398..1400
                     /label=L>CTG
     misc_feature    1425..1427
                     /label=L>CTG
     misc_feature    1437..1439
                     /label=P>CCG
     misc_feature    1452..1454
                     /label=P>CCG
     misc_feature    1461..1463
                     /label=G>GGC
     misc_feature    1485..1487
                     /label=P>CCG
     misc_feature    1491..1493
                     /label=P>CCG
     misc_feature    1503..1505
                     /label=S>TCT
     misc_feature    1506..1508
                     /label=P>CCG
     misc_RNA        1520
                     /label=NcoI delete C>G Ala site
     misc_feature    1536..1538
                     /label=G>GGC
     misc_feature    1554..1556
                     /label=T>ACC
     misc_feature    1581..1583
                     /label=L>CTG
     misc_feature    1611..1613
                     /label=G>GGC
     misc_feature    1626..1628
                     /label=G>GGC
     misc_feature    1644..1646
                     /label=G>GGC
     misc_feature    1656..1658
                     /label=P>CCG
     misc_feature    1668..1670
                     /label=P>CCG
     misc_feature    1683..1685
                     /label=S>TCT
     misc_feature    1707..1709
                     /label=R>CGU
     misc_feature    1710..1712
                     /label=S>TCT
     misc_feature    1725..1727
                     /label=P>CCG
     misc_feature    1746..1748
                     /label=S>AGC
     misc_feature    1761..1763
                     /label=S>AGC
     misc_feature    1773..1775
                     /label=G>GGC
     misc_feature    1827..1829
                     /label=P>CCG
     misc_feature    1884..1886
                     /label=L>CTG
     misc_feature    1905..1907
                     /label=L>CTG
     primer_bind     complement(1952..1972)
                     /label=nm74
     primer_bind     complement(1955..1972)
                     /label=mCherry nostop BamHI
     primer_bind     complement(1957..1972)
                     /label=nm86
     primer_bind     1967..1991
                     /label=nm235
     primer_bind     1969..1990
                     /label=nm39
     primer_bind     1972..1994
                     /label=XhoI-linker-mCherry
     primer_bind     1972..1991
                     /label=nm73
     CDS             1974..2690
                     /codon_start=1
                     /product="enhanced monomeric variant of CFP (Goedhart et 
                     al., 2012)"
                     /label=mTurquoise2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYFSDNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2675..2696)
                     /label=nm74
     primer_bind     complement(2678..2697)
                     /label=mCherry nostop BamHI
     primer_bind     complement(2680..2708)
                     /label=nm86
     misc_feature    2879..2998
                     /label=rRNA_
                     /label=rRNA_(rrnB)_1
     misc_feature    3002..3045
                     /label=rrnB_T1
     misc_feature    3177..3204
                     /label=rrnB_T2
     primer_bind     complement(3275..3302)
                     /label=T7
     primer_bind     complement(3301..3318)
                     /label=M13-fwd
     misc_feature    3313..3318
                     /label=-35_Signal
                     /label=-35_Signal(trp)_1(1)
     CDS             3389..3457
                     /label=LacZ alpha
     misc_feature    3896..4555
                     /label=Cm
                     /label=Cm(R)
     misc_feature    join(5668..5796,1..954)
                     /label=CDS
                     /label=CDS(lacIq)_2
ORIGIN
        1 gagctgaatt acattcccaa ccgcgtggca caacaactgg cgggcaaaca gtcgttgctg
       61 attggcgttg ccacctccag tctggccctg cacgcgccgt cgcaaattgt cgcggcgatt
      121 aaatctcgcg ccgatcaact gggtgccagc gtggtggtgt cgatggtaga acgaagcggc
      181 gtcgaagcct gtaaagcggc ggtgcacaat cttctcgcgc aacgcgtcag tgggctgatc
      241 attaactatc cgctggatga ccaggatgcc attgctgtgg aagctgcctg cactaatgtt
      301 ccggcgttat ttcttgatgt ctctgaccag acacccatca acagtattat tttctcccat
      361 gaagacggta cgcgactggg cgtggagcat ctggtcgcat tgggtcacca gcaaatcgcg
      421 ctgttagcgg gcccattaag ttctgtctcg gcgcgtctgc gtctggctgg ctggcataaa
      481 tatctcactc gcaatcaaat tcagccgata gcggaacggg aaggcgactg gagtgccatg
      541 tccggttttc aacaaaccat gcaaatgctg aatgagggca tcgttcccac tgcgatgctg
      601 gttgccaacg atcagatggc gctgggcgca atgcgcgcca ttaccgagtc cgggctgcgc
      661 gttggtgcgg atatctcggt agtgggatac gacgataccg aagacagctc atgttatatc
      721 ccgccgttaa ccaccatcaa acaggatttt cgcctgctgg ggcaaaccag cgtggaccgc
      781 ttgctgcaac tctctcaggg ccaggcggtg aagggcaatc agctgttgcc cgtctcactg
      841 gtgaaaagaa aaaccaccct ggcgcccaat acgcaaaccg cctctccccg cgcgttggcc
      901 gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa
      961 cgcaattaat gtgagttagc gcgaattgat ctggtttgac agcttatcat cgactgcacg
     1021 gtgcaccaat gcttctggcg tcaggcagcc atcggaagct gtggtatggc tgtgcaggtc
     1081 gtaaatcact gcataattcg tgtcgctcaa ggcgcactcc cgttctggat aatgtttttt
     1141 gcgccgacat cataacggtt ctggcaaata ttctgaaatg agctgtttac aattaatcat
     1201 ccggctcgta taatgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagacca
     1261 tggtgagcaa gggcgaggag gataacatgg cctctctccc agcgacacat gagttacaca
     1321 tctttggctc catcaacggt gtggactttg acatggtggg tcagggcacc ggcaatccaa
     1381 atgatggtta tgaggagtta aacctgaagt ccaccaaggg tgacctccag ttctccccct
     1441 ggattctggt ccctcatatc gggtatggct tccatcagta cctgccctac cctgacggga
     1501 tgtcgccttt ccaggccgcg atggtagatg gctccggata ccaagtccat cgcacaatgc
     1561 agtttgaaga tggtgcctcc cttactgtta actaccgcta cacctacgag ggaagccaca
     1621 tcaaaggaga ggcccaggtg aaggggactg gtttccctgc tgacggtcct gtgatgacca
     1681 actcgctgac cgctgcggac tggtgcaggt cgaagaagac ttaccccaac gacaaaacca
     1741 tcatcagtac ctttaagtgg agttacacca ctggaaatgg caagcgctac cggagcactg
     1801 cgcggaccac ctacaccttt gccaagccaa tggcggctaa ctatctgaag aaccagccga
     1861 tgtacgtgtt ccgtaagacg gagctcaagc actccaagac cgagctcaac ttcaaggagt
     1921 ggcaaaaggc ctttaccgat gtgatgggca tggacgagct gtacaaggaa ttgatggtga
     1981 gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg
     2041 taaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc tacggcaagc
     2101 tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc accctcgtga
     2161 ccaccctgtc ctggggcgtg cagtgcttcg cccgctaccc cgaccacatg aagcagcacg
     2221 acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc ttcttcaagg
     2281 acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc
     2341 gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg
     2401 agtacaacta ctttagcgac aacgtctata tcaccgccga caagcagaag aacggcatca
     2461 aggccaactt caagatccgc cacaacatcg aggacggcgg cgtgcagctc gccgaccact
     2521 accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac cactacctga
     2581 gcacccagtc caagctgagc aaagacccca acgagaagcg cgatcacatg gtcctgctgg
     2641 agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtacaag gaattctaaa
     2701 gcttgggccc gaacaaaaac tcatctcaga agaggatctg aatagcgccg tcgaccatca
     2761 tcatcatcat cattgagttt aaacggtctc cagcttggct gttttggcgg atgagagaag
     2821 attttcagcc tgatacagat taaatcagaa cgcagaagcg gtctgataaa acagaatttg
     2881 cctggcggca gtagcgcggt ggtcccacct gaccccatgc cgaactcaga agtgaaacgc
     2941 cgtagcgccg atggtagtgt ggggtctccc catgcgagag tagggaactg ccaggcatca
     3001 aataaaacga aaggctcagt cgaaagactg ggcctttcgt tttatctgtt gtttgtcggt
     3061 gaacgctctc ctgagtagga caaatccgcc gggagcggat ttgaacgttg cgaagcaacg
     3121 gcccggaggg tggcgggcag gacgcccgcc ataaactgcc aggcatcaaa ttaagcagaa
     3181 ggccatcctg acggatggcc tttttgcgtt tctacaaact cttttgttta tttttctaaa
     3241 tacattcaaa tatgtatccg ctcatgccaa ttcgccctat agtgagtcgt attacaattc
     3301 actggccgtc gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg
     3361 ccttgcagca catccccctt tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg
     3421 cccttcccaa cagttgcgca gcctgaatgg cgaatgggac gcgccctgta gcggcgcatt
     3481 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgctgatgt
     3541 ccggcggtgc ttttgccgtt acgcaccacc ccgtcagtag ctgaacagga gggacagctg
     3601 atagaaacag aagccactgg agcacctcaa aaacaccatc atacactaaa tcagtaagtt
     3661 ggcagcatca cccgacgcac tttgcgccga ataaatacct gtgacggaag atcacttcgc
     3721 agaataaata aatcctggtg tccctgttga taccgggaag ccctgggcca acttttggcg
     3781 aaaatgagac gttgatcggc acgtaagagg ttccaacttt caccataatg aaataagatc
     3841 actaccgggc gtattttttg agttatcgag attttcagga gctaaggaag ctaaaatgga
     3901 gaaaaaaatc actggatata ccaccgttga tatatcccaa tggcatcgta aagaacattt
     3961 tgaggcattt cagtcagttg ctcaatgtac ctataaccag accgttcagc tggatattac
     4021 ggccttttta aagaccgtaa agaaaaataa gcacaagttt tatccggcct ttattcacat
     4081 tcttgcccgc ctgatgaatg ctcatccgga atttcgtatg gcaatgaaag acggtgagct
     4141 ggtgatatgg gatagtgttc acccttgtta caccgttttc catgagcaaa ctgaaacgtt
     4201 ttcatcgctc tggagtgaat accacgacga tttccggcag tttctacaca tatattcgca
     4261 agatgtggcg tgttacggtg aaaacctggc ctatttccct aaagggttta ttgagaatat
     4321 gtttttcgtc tcagccaatc cctgggtgag tttcaccagt tttgatttaa acgtggccaa
     4381 tatggacaac ttcttcgccc ccgttttcac tatgggcaaa tattatacgc aaggcgacaa
     4441 ggtgctgatg ccgctggcga ttcaggttca tcatgccgtc tgtgatggct tccatgtcgg
     4501 cagaatgctt aatgaattac aacagtactg cgatgagtgg cagggcgggg cgtaattttt
     4561 ttaaggcagt tattggtgcc cttaaacgcc tggtgctacg cctgaataag tgataataag
     4621 cggatgaatg gcagaaattc gaaggttctc tgagctacca actctttgaa ccgaggtaac
     4681 tggcttggag gagcgcagtc accaaaactt gtcctttcag tttagcctta accggcgcat
     4741 gacttcaaga ctaactcctc taaatcaatt accagtggct gctgccagtg gtgcttttgc
     4801 atgtctttcc gggttggact caagacgata gttaccggat aaggcgcagc ggtcggactg
     4861 aacggggggt tcgtgcatac agtccagctt ggagcgaact gcctacccgg aactgagtgt
     4921 caggcgtgga atgagacaaa cgcggccata acagcggaat gacaccggta aaccgaaagg
     4981 caggaacagg agagcgcacg agggagccgc cagggggaaa cgcctggtat ctttatagtc
     5041 ctgtcgggtt tcgccaccac tgatttgagc gtcagatttc gtgatgcttg tcaggggggc
     5101 ggagcctatg gaaaaacggc tttgccgcgg ccctctcact tccctgttaa gtatcttcct
     5161 ggcatcttcc aggaaatctc cgccccgttc gtaagccatt tccgctcgcc gcagtcgaac
     5221 gaccgagcgt agcgagtcag tgagcgagga agcggaatat atcctgtatc acatattctg
     5281 ctgacgcacc ggtgcagcct tttttctcct gccacatgaa gcacttcact gacaccctca
     5341 tcagtgccaa catagtaagc cagtatacac tccgctatcg ctacgtgact gggtcatggc
     5401 tgcgccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc
     5461 atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc
     5521 gtcatcaccg aaacgcgcga ggcagcagat caattcgcgc gcgaaggcga agcggcatgc
     5581 atttacgttg acaccatcga atggtgcaaa acctttcgcg gtatggcatg atagcgcccg
     5641 gaagagagtc aattcagggt ggtgaatgtg aaaccagtaa cgttatacga tgtcgcagag
     5701 tatgccggtg tctcttatca gaccgtttcc cgcgtggtga accaggccag ccacgtttct
     5761 gcgaaaacgc gggaaaaagt ggaagcggcg atggcg
//
