LOCUS       Exported                6227 bp ds-DNA     circular SYN 24-FEB-2019
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6227)
  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..6227
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        53..81
                     /label=TrcDown
     misc_feature    53..58
                     /label=TTTACA (-35)
     misc_feature    76..81
                     /label=TATAAT (-10)
     misc_feature    114..118
                     /label=RBS (CAGGA)
     CDS             129..787
                     /label=OmpA
     primer_bind     784..813
                     /label=nm75
     primer_bind     784..813
                     /label=XhoI-linker-mCherry
     primer_bind     787..810
                     /label=nm235
     primer_bind     788..809
                     /label=nm39
     CDS             793..1500
                     /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     793..813
                     /label=age1-mapple
     primer_bind     793..810
                     /label=nm234 ncoi-fp f
     misc_feature    829..831
                     /label=L>CTG
     misc_feature    832..834
                     /label=P>CCG
     misc_feature    838..840
                     /label=T>ACC
     misc_feature    847..849
                     /label=L>CTG
     misc_feature    910..912
                     /label=P>CCG
     misc_feature    931..933
                     /label=L>CTG
     misc_feature    958..960
                     /label=L>CTG
     misc_feature    970..972
                     /label=P>CCG
     misc_feature    985..987
                     /label=P>CCG
     misc_feature    994..996
                     /label=G>GGC
     misc_feature    1018..1020
                     /label=P>CCG
     misc_feature    1024..1026
                     /label=P>CCG
     misc_feature    1036..1038
                     /label=S>TCT
     misc_feature    1039..1041
                     /label=P>CCG
     misc_RNA        1053
                     /label=NcoI delete C>G Ala site
     misc_feature    1069..1071
                     /label=G>GGC
     misc_feature    1087..1089
                     /label=T>ACC
     misc_feature    1114..1116
                     /label=L>CTG
     misc_feature    1144..1146
                     /label=G>GGC
     misc_feature    1159..1161
                     /label=G>GGC
     misc_feature    1177..1179
                     /label=G>GGC
     misc_feature    1189..1191
                     /label=P>CCG
     misc_feature    1201..1203
                     /label=P>CCG
     misc_feature    1216..1218
                     /label=S>TCT
     misc_feature    1240..1242
                     /label=R>CGU
     misc_feature    1243..1245
                     /label=S>TCT
     misc_feature    1258..1260
                     /label=P>CCG
     misc_feature    1279..1281
                     /label=S>AGC
     misc_feature    1294..1296
                     /label=S>AGC
     misc_feature    1306..1308
                     /label=G>GGC
     misc_feature    1360..1362
                     /label=P>CCG
     misc_feature    1417..1419
                     /label=L>CTG
     misc_feature    1438..1440
                     /label=L>CTG
     primer_bind     complement(1485..1505)
                     /label=nm74
     primer_bind     complement(1488..1505)
                     /label=mCherry nostop BamHI
     primer_bind     complement(1490..1505)
                     /label=nm86
     primer_bind     1500..1524
                     /label=nm235
     primer_bind     1502..1523
                     /label=nm39
     primer_bind     1505..1527
                     /label=nm75
     primer_bind     1505..1527
                     /label=XhoI-linker-mCherry
     misc_feature    1507..2223
                     /label=sfTurquoise2
     primer_bind     1507..1527
                     /label=age1-mapple
     primer_bind     1507..1524
                     /label=nm234 ncoi-fp f
     misc_feature    1597..1599
                     /label=S30R
     misc_feature    1624..1626
                     /label=Y39N
     primer_bind     1633..1674
                     /label=Primer 2
     primer_bind     complement(1633..1674)
                     /label=nm227
     primer_bind     1633..1674
                     /label=nm226
     variation       1651..1653
                     /label=C48
     primer_bind     1695..1734
                     /label=Primer 3
     primer_bind     complement(1695..1734)
                     /label=Primer 1
     primer_bind     1695..1734
                     /label=nm228
     variation       1719
                     /label=C70
     misc_feature    1804..1806
                     /label=F99S
     misc_feature    1822..1824
                     /label=N105T
     misc_feature    2020..2022
                     /label=I171V
     primer_bind     complement(2208..2229)
                     /label=nm74
     primer_bind     complement(2211..2230)
                     /label=mCherry nostop BamHI
     primer_bind     complement(2213..2238)
                     /label=nm86
     CDS             2757..3615
                     /label=AmpR
     CDS             5089..6046
                     /label=LacIq
ORIGIN
        1 gttttttgcg ccgacatcat aacggttctg gcaaatattc tgaaatgagc tgtttacaat
       61 taatcatccg gctcgtataa tgtgtggaat tgtgagcgga taacaatttc acacaggaaa
      121 cagaccatgg caaaaaagac agctatcgcg attgcagtgg cactggctgg tttcgctacc
      181 gtagcgcagg ccgctccgaa agataacacc tggtacactg gtgctaaact gggctggtcc
      241 cagtaccatg acactggcct catcggccaa cgcttggaga tctgccagaa cgtttgctac
      301 tacctcggga cgctcaatgg tccgacccat gaaaaccaac tgggcgctgg tgcttttggt
      361 ggttaccagg ttaacccgta tgttggcttt gaaatgggtt acgactggtt aggtcgtatg
      421 ccgtacaaag gcagcgttga aaacggtgca tacaaagctc agggcgttca actgaccgct
      481 aaactgggtt acccaatcac tgacgacctg gacatctaca ctcgtctggg tggcatggta
      541 tggcgtgcag acactaaatc caacgtttat ggtaaaaacc acgacaccgg cgtttctccg
      601 gtcttcgctg gcggtgttga gtacgcgatc actcctgaaa tcgctacccg tctggaatac
      661 cagtggacca acaacatcgg tgacgcaggc cagatcggcc agcgtccgga caacggcatg
      721 ctgagcctgg gtgtttccta ccgtttcggt cagggcgaag cagctccact tgaggatccc
      781 cccgctgaat tgatggtgag caagggcgag gaggataaca tggcctctct cccagcgaca
      841 catgagttac acatctttgg ctccatcaac ggtgtggact ttgacatggt gggtcagggc
      901 accggcaatc caaatgatgg ttatgaggag ttaaacctga agtccaccaa gggtgacctc
      961 cagttctccc cctggattct ggtccctcat atcgggtatg gcttccatca gtacctgccc
     1021 taccctgacg ggatgtcgcc tttccaggcc gcgatggtag atggctccgg ataccaagtc
     1081 catcgcacaa tgcagtttga agatggtgcc tcccttactg ttaactaccg ctacacctac
     1141 gagggaagcc acatcaaagg agaggcccag gtgaagggga ctggtttccc tgctgacggt
     1201 cctgtgatga ccaactcgct gaccgctgcg gactggtgca ggtcgaagaa gacttacccc
     1261 aacgacaaaa ccatcatcag tacctttaag tggagttaca ccactggaaa tggcaagcgc
     1321 taccggagca ctgcgcggac cacctacacc tttgccaagc caatggcggc taactatctg
     1381 aagaaccagc cgatgtacgt gttccgtaag acggagctca agcactccaa gaccgagctc
     1441 aacttcaagg agtggcaaaa ggcctttacc gatgtgatgg gcatggacga gctgtacaag
     1501 gaattgatgg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag
     1561 ctggacggcg acgtaaacgg ccacaagttc agcgtgcgcg gcgagggcga gggcgatgcc
     1621 accaatggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg
     1681 cccaccctcg tgaccaccct gtcctggggt gtacaggtct tcgcccgcta ccccgaccac
     1741 atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc
     1801 atcagcttca aggacgacgg cacatacaag acccgcgccg aggtgaagtt cgagggcgac
     1861 accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg
     1921 gggcacaagc tggagtacaa ctactttagc gacaacgtct atatcaccgc cgacaagcag
     1981 aagaacggca tcaaggccaa cttcaagatc cgccacaacg tggaggacgg cggcgtgcag
     2041 ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac
     2101 aaccactacc tgagcaccca gtccaagctg agcaaagacc ccaacgagaa gcgcgatcac
     2161 atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac
     2221 aaggaattct aaagcttggc tgttttggcg gatgagagaa gattttcagc ctgatacaga
     2281 ttaaatcaga acgcagaagc ggtctgataa aacagaattt gcctggcggc agtagcgcgg
     2341 tggtcccacc tgaccccatg ccgaactcag aagtgaaacg ccgtagcgcc gatggtagtg
     2401 tggggtctcc ccatgcgaga gtagggaact gccaggcatc aaataaaacg aaaggctcag
     2461 tcgaaagact gggcctttcg ttttatctgt tgtttgtcgg tgaacgctct cctgagtagg
     2521 acaaatccgc cgggagcgga tttgaacgtt gcgaagcaac ggcccggagg gtggcgggca
     2581 ggacgcccgc cataaactgc caggcatcaa attaagcaga aggccatcct gacggatggc
     2641 ctttttgcgt ttctacaaac tctttttgtt tatttttcta aatacattca aatatgtatc
     2701 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
     2761 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     2821 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
     2881 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     2941 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgtg
     3001 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     3061 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     3121 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
     3181 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     3241 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgccta
     3301 cagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
     3361 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     3421 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
     3481 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     3541 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     3601 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
     3661 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     3721 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     3781 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     3841 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     3901 ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc
     3961 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     4021 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     4081 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     4141 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     4201 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     4261 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     4321 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     4381 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     4441 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata
     4501 ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc
     4561 gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atatggtgca
     4621 ctctcagtac aatctgctct gatgccgcat agttaagcca gtatacactc cgctatcgct
     4681 acgtgactgg gtcatggctg cgccccgaca cccgccaaca cccgctgacg cgccctgacg
     4741 ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat
     4801 gtgtcagagg ttttcaccgt catcaccgaa acgcgcgagg cagcagatca attcgcgcgc
     4861 gaaggcgaag cggcatgcat ttacgttgac accatcgaat ggtgcaaaac ctttcgcggt
     4921 atggcatgat agcgcccgga agagagtcaa ttcagggtgg tgaatgtgaa accagtaacg
     4981 ttatacgatg tcgcagagta tgccggtgtc tcttatcaga ccgtttcccg cgtggtgaac
     5041 caggccagcc acgtttctgc gaaaacgcgg gaaaaagtgg aagcggcgat ggcggagctg
     5101 aattacattc ccaaccgcgt ggcacaacaa ctggcgggca aacagtcgtt gctgattggc
     5161 gttgccacct ccagtctggc cctgcacgcg ccgtcgcaaa ttgtcgcggc gattaaatct
     5221 cgcgccgatc aactgggtgc cagcgtggtg gtgtcgatgg tagaacgaag cggcgtcgaa
     5281 gcctgtaaag cggcggtgca caatcttctc gcgcaacgcg tcagtgggct gatcattaac
     5341 tatccgctgg atgaccagga tgccattgct gtggaagctg cctgcactaa tgttccggcg
     5401 ttatttcttg atgtctctga ccagacaccc atcaacagta ttattttctc ccatgaagac
     5461 ggtacgcgac tgggcgtgga gcatctggtc gcattgggtc accagcaaat cgcgctgtta
     5521 gcgggcccat taagttctgt ctcggcgcgt ctgcgtctgg ctggctggca taaatatctc
     5581 actcgcaatc aaattcagcc gatagcggaa cgggaaggcg actggagtgc catgtccggt
     5641 tttcaacaaa ccatgcaaat gctgaatgag ggcatcgttc ccactgcgat gctggttgcc
     5701 aacgatcaga tggcgctggg cgcaatgcgc gccattaccg agtccgggct gcgcgttggt
     5761 gcggatatct cggtagtggg atacgacgat accgaagaca gctcatgtta tatcccgccg
     5821 ttaaccacca tcaaacagga ttttcgcctg ctggggcaaa ccagcgtgga ccgcttgctg
     5881 caactctctc agggccaggc ggtgaagggc aatcagctgt tgcccgtctc actggtgaaa
     5941 agaaaaacca ccctggcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca
     6001 ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat
     6061 taatgtgagt tagcgcgaat tgatctggtt tgacagctta tcatcgactg cacggtgcac
     6121 caatgcttct ggcgtcaggc agccatcgga agctgtggta tggctgtgca ggtcgtaaat
     6181 cactgcataa ttcgtgtcgc tcaaggcgca ctcccgttct ggataat
//
