LOCUS       Exported                3980 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Expresses sfYFP under Cin promoter.
ACCESSION   .
VERSION     .
KEYWORDS    C309
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3980)
  AUTHORS   Alnahhas RN, Sadeghpour M, Chen Y, Frey AA, Ott W, Josic K, Bennett 
            MR
  TITLE     Majority sensing in synthetic microbial consortia.
  JOURNAL   Nat Commun. 2020 Jul 21;11(1):3659. doi: 10.1038/s41467-020-17475-z.
  PUBMED    32694598
REFERENCE   2  (bases 1 to 3980)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..3980
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     protein_bind    58..74
                     /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)."
     primer_bind     63..85
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     CDS             275..304
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     protein_bind    458..476
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    488..506
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     CDS             590..1303
                     /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
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     terminator      1489..1516
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(1665..2480)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSSMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     2388..2407
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2472..2491
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      2602..3190
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3091..3110
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3344..3361
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3376..3515
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3441..3460)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3601..3623
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3617..3808)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 agcttcccaa ccttaccaga gggcgcccca gctggcaatt ccgacgtctg tgtggaattg
       61 tgagcggata acaatttcac acagggccct cggacaccga ggagaatgtc aagaggcgaa
      121 cacacaacgt cttggagcgc cagaggagga acgagctaaa acggagcttt ttgccctgcg
      181 tgaccagatc ccggagttgg aaaacaatga aaaggccccc aaggtagtta tccttaaaaa
      241 agccacagca tacatcctgt ccgtccaagc agaggagcaa aagctcattt ctgaagagga
      301 cttgttgcgg aaacgacgag aacagttgaa acacaaactt gaacagctac ggaactcttg
      361 tgcgtggcgc atgtcgttat cgcaaaaccg ctgcacactt ttgcgcgaca tgctctgatc
      421 cccctcatct gggggggcct atctgaggga atttacttcc ctatcagtga tagagataat
      481 gagcacatcc ctatcagtga tagagatgca tgccacttaa aaaggagatc aacaatgaaa
      541 gcaattttcg tactgaaaca tcttaatcat gctaaggaga aatactagta tgcgtaaagg
      601 cgaagagctg ttcactggtg tcgtccctat tctggtggaa ctggatggtg atgtcaacgg
      661 tcataagttt tccgtgcgtg gcgagggtga aggtgacgca actaatggta aactgacgct
      721 gaagttcatc tgtactactg gtaaactgcc ggtaccttgg ccgactctgg taacgacgct
      781 gacttatggt gttcagtgct ttgctcgtta tccggaccat atgaagcagc atgacttctt
      841 caagtccgcc atgccggaag gctatgtgca ggaacgcacg atttccttta aggatgacgg
      901 cacgtacaaa acgcgtgcgg aagtgaaatt tgaaggcgat accctggtaa accgcattga
      961 gctgaaaggc attgacttta aagaagacgg caatatcctg ggccataagc tggaatacaa
     1021 ttttaacagc cacaatgttt acatcaccgc cgataaacaa aaaaatggca ttaaagcgaa
     1081 ttttaaaatt cgccacaacg tggaggatgg cagcgtgcag ctggctgatc actaccagca
     1141 aaacactcca atcggtgatg gtcctgttct gctgccagac aatcactatc tgagctacca
     1201 aagcgttctg tctaaagatc cgaacgagaa acgcgatcat atggttctgc tggagttcgt
     1261 aaccgcagcg ggcatcacgc atggtatgga tgaactgtac aaagcagcga acgacgaaaa
     1321 ttacgccctt gcagcgtaat aaaagcttcc cctacgtgct gctgaagttg cccgcaacag
     1381 agagtggaac caaccggtga taccacgata ctatgactga gagtcaacgc catgagcggc
     1441 ctcatttctt attctgagtt acaacagtcc gcaccgctgt ctcacactgg ctcaccttcg
     1501 ggtgggcctt tctgcgttta tatactagag agagaatata aaaagccaga ttattaatcc
     1561 ggctttttta ttatttccta gctagccggg gaaatgtgcg cggaacccct atttgtttat
     1621 ttttctaaat acattcaaat atgtatccgc tcatgaatta attcttagaa aaactcatcg
     1681 agcatcaaat gaaactgcaa tttattcata tcaggattat caataccata tttttgaaaa
     1741 agccgtttct gtaatgaagg agaaaactca ccgaggcagt tccataggat ggcaagatcc
     1801 tggtatcggt ctgcgattcc gactcgtcca acatcaatac aacctattaa tttcccctcg
     1861 tcaaaaataa ggttatcaag tgagaaatca ccatgagtga cgactgaatc cggtgagaat
     1921 ggcaaaagtt tatgcatttc tttccagact tgttcaacag gccagccatt acgctcgtca
     1981 tcaaaatcac tcgcatcaac caaaccgtta ttcattcgtg attgcgcctg agcgagacga
     2041 aatacgcgat cgctgttaaa aggacaatta caaacaggaa tcgaatgcaa ccggcgcagg
     2101 aacactgcca gcgcatcaac aatattttca cctgaatcag gatattcttc taatacctgg
     2161 aatgctgttt tcccggggat cgcagtggtg agtaaccatg catcatcagg agtacggata
     2221 aaatgcttga tggtcggaag aggcataaat tccgtcagcc agtttagtct gaccatctca
     2281 tctgtaacat cattggcaac gctacctttg ccatgtttca gaaacaactc tggcgcatcg
     2341 ggcttcccat acaatcgata gattgtcgca cctgattgcc cgacattatc gcgagcccat
     2401 ttatacccat ataaatcagc atccatgctg gaatttaatc gcggcctaga gcaagacgtt
     2461 tcccgttgaa tatggctcat aacacccctt gtattactgt ttatgtaagc agacagtttt
     2521 attgttcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     2581 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     2641 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     2701 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc ttctagtgta
     2761 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     2821 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     2881 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     2941 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     3001 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     3061 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     3121 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     3181 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     3241 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
     3301 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
     3361 ggaagcggaa gagcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca
     3421 ccgcaatggt gcactctcag tacaatctgc tctgatgccg catagttaag ccagtataca
     3481 ctccgctatc gctacgtgac tgggtcatgg ctgcgccccg acacccgcca acacccgctg
     3541 acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct
     3601 ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg aggcagctgc
     3661 ggtaaagctc atcagcgtgg tcgtgaagcg attcacagat gtctgcctgt tcatccgcgt
     3721 ccagctcgtt gagtttctcc agaagcgtta atgtctggct tctgataaag cgggccatgt
     3781 taagggcggt tttttcctgt ttggtcactg atgcctccgt gtaaggggga tttctgttca
     3841 tgggggtaat gataccgatg aaacgagaga ggatgctcac gatacgggtt actgatgatg
     3901 aacatgcccg gttactggaa cgttgtgagg gtaaacaact ggcggtatgg atgcggcggg
     3961 accagagctc agtgcgctcg
//
