LOCUS       Exported                2606 bp ds-DNA     circular SYN 13-JUN-2018
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2606)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 3, 2018 from SnapGene 4.2.6
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..2606
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     misc_feature    1..960
                     /label=from pVRb32_1122
     promoter        1..80
                     /label=Pecf32_1122 (-60 to +20)
     primer_bind     complement(43..64)
                     /label=pECF32TSS_R1
     misc_feature    81..86
                     /label=BamHI site
     misc_feature    87..115
                     /label=strong synthetic RBS
                     /note="\cite{Segall-Shapiro2014}"
     CDS             116..835
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK*"
     primer_bind     640..651
                     /label=21_2
     terminator      874..960
                     /label=rrnB T1 terminator
     primer_bind     complement(906..933)
                     /label=GFP_R
     primer_bind     909..945
                     /label=ColE1_F
     primer_bind     complement(1039..1078)
                     /label=repColE2_R1
     primer_bind     1039..1078
                     /label=repColE2_F2
     misc_feature    1121..1589
                     /label=Ori of ColE2
     primer_bind     1204..1220
                     /label=repColE2_seq3
     primer_bind     complement(1646..1685)
                     /label=repColE2_R2
     primer_bind     1646..1685
                     /label=repColE2_F3
     primer_bind     complement(1670..1711)
                     /label=Sp_R1
     primer_bind     1670..1711
                     /label=Sp_F1
     terminator      1703..1797
                     /label=lambda t0 terminator
     primer_bind     complement(1814..1840)
                     /label=ColE1_R
     primer_bind     1816..1843
                     /label=Cm_F
     CDS             complement(1828..2487)
                     /codon_start=1
                     /label=CmR
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     1869..1885
                     /label=repColE2_seq4
     primer_bind     2016..2033
                     /label=21_Cm_4
     primer_bind     2534..2553
                     /label=recColE2_seq1
     primer_bind     complement(join(2586..2606,1..25))
                     /label=HH02_32_R
     primer_bind     join(2586..2606,1..25)
                     /label=HH02_32_F
ORIGIN
        1 gcgactttat ttaacagcgg catggccagg gaaccgatgc gtcaatcgca ccacacaatg
       61 acaactgctc tcatcattga ggatcctgtc aatttccgcg atagaggagg taaagatgag
      121 caaaggagaa gaacttttca ctggagttgt cccaattctt gttgaattag atggtgatgt
      181 taatgggcac aaattttctg tccgtggaga gggtgaaggt gatgctacaa acggaaaact
      241 cacccttaaa tttatttgca ctactggaaa actacctgtt ccgtggccaa cacttgtcac
      301 tactctgacc tatggtgttc aatgcttttc ccgttatccg gatcacatga aacggcatga
      361 ctttttcaag agtgccatgc ccgaaggtta tgtacaggaa cgcactatat ctttcaaaga
      421 tgacgggacc tacaagacgc gtgctgaagt caagtttgaa ggtgataccc ttgttaatcg
      481 tatcgagtta aagggtattg attttaaaga agatggaaac attcttggac acaaactcga
      541 gtacaacttt aactcacaca atgtatacat cacggcagac aaacaaaaga atggaatcaa
      601 agctaacttc aaaattcgcc acaacgttga agatggttcc gttcaactag cagaccatta
      661 tcaacaaaat actccaattg gcgatggccc tgtcctttta ccagacaacc attacctgtc
      721 gacacaatct gtcctttcga aagatcccaa cgaaaagcgt gaccacatgg tccttcttga
      781 gtttgtaact gctgctggga ttacacatgg catggatgag ctctacaaat aataaatgtc
      841 cagacctgca ggcatgcaag cctctagagg catcaaataa aacgaaaggc tcagtcgaaa
      901 gactgggcct ttcgttttat ctgttgtttg tcggtgaacg ctctcctgag taggacaaat
      961 ccgccgccct agacctaggg cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     1021 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     1081 cagcaaaagg ccaggaaccg taaaaaggcc gccctcgagg agcgcctcag cgcgccgtag
     1141 cgtcgataaa aattacgggc tggggcgaaa ctaccatctg ttcgaaaagg tccgtaaatg
     1201 ggcctacaga gcgattcgtc agggctggcc tgtattctca caatggcttg atgccgttat
     1261 ccagcgtgtc gaaatgtaca acgcttcgct tcccgttccg ctttctccgg ctgaatgtcg
     1321 ggctattggc aagagcattg cgaaatatac acacaggaaa ttctcaccag agggattttc
     1381 cgctgtacag gccgctcgcg gtcgcaaggg cggaactaaa tctaagcgcg cagcagttcc
     1441 tacatcagca cgttcgctga aaccgtggga ggcattaggc atcagtcgag cgacgtacta
     1501 ccgaaaatta aaatgtgacc cagacctcgc aaaatgagac cagataagcc ttatcagata
     1561 acagcgccct tttggcgtct ttttgagcac ctaggtgtaa aacgacggcc agtcttaagc
     1621 tcgggccctc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     1681 tttggtcatg actagtgctt ggattctcac caataaaaaa cgcccggcgg caaccgagcg
     1741 ttctgaacaa atccagatgg agttctgagg tcattactgg atctatcaac aggagtccaa
     1801 gcgagctctc gaaccccaga gtcccgctta cgccccgccc tgccactcat cgcagtactg
     1861 ttgtaattca ttaagcattc tgccgacatg gaagccatca caaacggcat gatgaacctg
     1921 aatcgccagc ggcatcagca ccttgtcgcc ttgcgtataa tatttgccca tggtgaaaac
     1981 gggggcgaag aagttgtcca tattggccac gtttaaatca aaactggtga aactcaccca
     2041 gggattggct gagacgaaaa acatattctc aataaaccct ttagggaaat aggccaggtt
     2101 ttcaccgtaa cacgccacat cttgcgaata tatgtgtaga aactgccgga aatcgtcgtg
     2161 gtattcactc cagagcgatg aaaacgtttc agtttgctca tggaaaacgg tgtaacaagg
     2221 gtgaacacta tcccatatca ccagctcacc gtctttcatt gccatacgaa attccggatg
     2281 agcattcatc aggcgggcaa gaatgtgaat aaaggccgga taaaacttgt gcttattttt
     2341 ctttacggtc tttaaaaagg ccgtaatatc cagctgaacg gtctggttat aggtacattg
     2401 agcaactgac tgaaatgcct caaaatgttc tttacgatgc cattgggata tatcaacggt
     2461 ggtatatcca gtgatttttt tctccatttt agcttcctta gctcctgaaa atctcgataa
     2521 ctcaaaaaat acgcccggta gtgatcttat ttcattatgg tgaaagttgg aacctcttac
     2581 gtgcccgatc aactcgagtg ccacct
//
