LOCUS       Exported                3987 bp DNA     circular SYN 10-DEC-2021
DEFINITION  ~out pTet GFP10-link-FRB/IRBS/FKBP link-GFP11.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3987)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Feb 1, 2022 from SnapGene 6.0.0
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..3987
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        6..79
                     /label=PLtetO-1 promoter
                     /note="modified phage lambda PL promoter with tet operator 
                     sites (Lutz and Bujard, 1997)"
                     /note="This forward directional feature has 5 segments:
                        1:  6 .. 24 / #ffffff
                        2: 25 .. 30 / #ffffff / -35
                        3: 31 .. 47 / #ffffff
                        4: 48 .. 53 / #ffffff / -10
                        5: 54 .. 79 / #ffffff"
     protein_bind    6..24
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
                     /note="color: #31849b"
     protein_bind    31..49
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
                     /note="color: #31849b"
     RBS             89..100
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
                     /note="color: #a6acb3"
     misc_feature    108..215
                     /label=GFP10-linker
                     /note="color: #a6acb3; direction: RIGHT"
     CDS             240..518
                     /codon_start=1
                     /product="FKBP-rapamycin binding domain of human FRAP"
                     /label=FRB
                     /note="color: #993366"
                     /translation="ILWHEMWHEGLEEASRLYFGERNVKGMFEVLEPLHAMMERGPQTL
                     KETSFNQAYGRDLMEAQEWCRKYMKSGNVKDLTQAWDLYYHVFRRISK"
     misc_feature    558..893
                     /label=FKBP
                     /note="color: #ff6600; direction: RIGHT"
     misc_feature    894..1001
                     /label=linker-GFP11
                     /note="color: #a6acb3; direction: RIGHT"
     misc_feature    1002..1004
                     /label=*
                     /note="color: #ffffff"
     CDS             1011..1028
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /note="color: #cc99b2"
                     /translation="HHHHHH"
     terminator      1117..1203
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
                     /note="color: #ffffff"
     rep_origin      complement(1367..1955)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #ffff00"
     terminator      2043..2137
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
                     /note="color: #ffffff"
     CDS             complement(2143..2766)
                     /codon_start=1
                     /gene="tetR from transposon Tn10"
                     /product="tetracycline repressor TetR"
                     /label=TetR
                     /note="TetR binds to the tetracycline operator tetO to 
                     inhibit transcription. This inhibition can be relieved by 
                     adding tetracycline or doxycycline."
                     /note="color: #993366"
                     /translation="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDHQGAEPAFLFGLELIIYGLEKQLKCESGS"
     CDS             complement(2807..3598)
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /note="color: #ccffcc"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
ORIGIN
        1 tcgagtccct atcagtgata gagattgaca tccctatcag tgatagagat actgagcaca
       61 tcagcaggac gcactgaccg agttcattaa agaggagaaa gatacccatg ggcgatttac
      121 cagacgacca ttacctgtcg acacaaacta tcctttcgaa agatctcaac ggaactgacg
      181 ttgggtctgg cggtggttcc ggtggtggag gttctcatat ggagctgatc cgagtggcca
      241 tcctctggca tgagatgtgg catgaaggcc tggaagaggc atctcgtttg tactttgggg
      301 aaaggaacgt gaaaggcatg tttgaggtgc tggagccctt gcatgctatg atggaacggg
      361 gcccccagac tctgaaggaa acatccttta atcaggccta tggtcgagat ttaatggagg
      421 cccaagagtg gtgcaggaag tacatgaaat cagggaatgt caaggacctc acccaagcct
      481 gggacctcta ttatcatgtg ttccgacgaa tctcaaagca gtaaggtacc taactaagta
      541 attaaaggaa acagacgatg actagtggag tgcaggtgga aaccatctcc ccaggaaacg
      601 ggcgcacctt ccccaagcgc ggccagacct gcgtggtgca ctacaccggg atgcttgaag
      661 atggaaagaa atttgattcc tcccgggaca gaaacaagcc ctttaagttt atgctaggca
      721 agcaggaggt gatccgaggc tgggaagaag gggttgccca gatgagtgtg ggtcagagag
      781 ccaaactgac tatatcacct gactatgcat acggagctac aggtcatcct ggaattattc
      841 cacctcacgc tactctcgtc ttcgatgtgg agcttctaaa actggaagga tccggtggcg
      901 gtggttcagg tggagggtct ggtggcggat caacatctga aaagcgtgac cacatggtcc
      961 ttcttgagta tgtaactgct gctgggatta cagatgcatc ttaactcgag caccaccacc
     1021 accaccactg agatccggct gctaacaaag cccgaaagga agctgagttg gctgctgcca
     1081 ccgctgagca ataactagca taacctctag aggcatcaaa taaaacgaaa ggctcagtcg
     1141 aaagactggg cctttcgttt tatctgttgt ttgtcggtga acgctctcct gagtaggaca
     1201 aatccgccgc cctagaccta ggcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     1261 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     1321 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     1381 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     1441 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     1501 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     1561 aatgctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     1621 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     1681 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     1741 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     1801 ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     1861 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     1921 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     1981 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg actagcgctt
     2041 ggattctcac caataaaaaa cgcccggcgg caaccgagcg ttctgaacaa atccagatgg
     2101 agttctgagg tcattactgg atctatcaac aggagtccaa gcttaagacc cactttcaca
     2161 tttaagttgt ttttctaatc cgtatatgat caattcaagg ccgaataaga aggctggctc
     2221 tgcaccttgg tgatcaaata attcgatagc ttgtcgtaat aatggcggca tactatcagt
     2281 agtaggtgtt tccctttctt ctttagcgac ttgatgctct tgatcttcca atacgcaacc
     2341 taaagtaaaa tgccccacag cgctgagtgc atataatgca ttctctagtg aaaaaccttg
     2401 ttggcataaa aaggctaatt gattttcgag agtttcatac tgtttttctg taggccgtgt
     2461 acctaaatgt acttttgctc catcgcgatg acttagtaaa gcacatctaa aacttttagc
     2521 gttattacgt aaaaaatctt gccagctttc cccttctaaa gggcaaaagt gagtatggtg
     2581 cctatctaac atctcaatgg ctaaggcgtc gagcaaagcc cgcttatttt ttacatgcca
     2641 atacaatgta ggctgctcta cacctagctt ctgggcgagt ttacgggttg ttaaaccttc
     2701 gattccgacc tcattaagca gctctaatgc gctgttaatc actttacttt tatctaatct
     2761 ggacatcatt aatgtttatt gagctctcga accccagagt cccgcattat ttgccgacta
     2821 ccttggtgat ctcgcctttc acgtagtgga caaattcttc caactgatct gcgcgcgagg
     2881 ccaagcgatc ttcttcttgt ccaagataag cctgtctagc ttcaagtatg acgggctgat
     2941 actgggccgg caggcgctcc attgcccagt cggcagcgac atccttcggc gcgattttgc
     3001 cggttactgc gctgtaccaa atgcgggaca acgtaagcac tacatttcgc tcatcgccag
     3061 cccagtcggg cggcgagttc catagcgtta aggtttcatt tagcgcctca aatagatcct
     3121 gttcaggaac cggatcaaag agttcctccg ccgctggacc taccaaggca acgctatgtt
     3181 ctcttgcttt tgtcagcaag atagccagat caatgtcgat cgtggctggc tcgaagatac
     3241 ctgcaagaat gtcattgcgc tgccattctc caaattgcag ttcgcgctta gctggataac
     3301 gccacggaat gatgtcgtcg tgcacaacaa tggtgacttc tacagcgcgg agaatctcgc
     3361 tctctccagg ggaagccgaa gtttccaaaa ggtcgttgat caaagctcgc cgcgttgttt
     3421 catcaagcct tacggtcacc gtaaccagca aatcaatatc actgtgtggc ttcaggccgc
     3481 catccactgc ggagccgtac aaatgtacgg ccagcaacgt cggttcgaga tggcgctcga
     3541 tgacgccaac tacctctgat agttgagtcg atacttcggc gatcaccgct tccctcatga
     3601 tgtttaactt tgttttaggg cgactgccct gctgcgtaac atcgttgctg ctccataaca
     3661 tcaaacatcg acccacggcg taacgcgctt gctgcttgga tgcccgaggc atagactgta
     3721 ccccaaaaaa acatgtcata acaagccatg aaaaccgcca ctgcgccgtt accatgcgaa
     3781 acgatcctca tcctgtctct tgatcagatc ttgatcccct gcgccatcag atccttggcg
     3841 gcaagaaagc catccagttt actttgcagg gcttcccaac cttaccagag ggcgccccag
     3901 ctggcaattc cgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc
     3961 gtatcacgag gccctttcgt cttcacc
//
