LOCUS       sequence-459776-        2967 bp    DNA     circular SYN 15-APR-2025
DEFINITION  lvl 0 5 gene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2967)
  AUTHORS   Duportet X, Wroblewska L, Guye P, Li Y, Eyquem J, Rieders J, 
            Rimchala T, Batt G, Weiss R
  TITLE     A platform for rapid prototyping of synthetic gene networks in 
            mammalian cells.
  JOURNAL   Nucleic Acids Res. 2014 Dec 1;42(21):13440-51. doi: 
            10.1093/nar/gku1082. Epub 2014 Nov 5.
  PUBMED    25378321
REFERENCE   2  (bases 1 to 2967)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1093/nar/gku1082"; journalName: "Nucleic Acids Res"; date: 
            "2014-12-1- 1"; volume: "42"; issue: "21"; pages: "13440-51"
COMMENT     Sequence Label: lvl0-G-rtTA
FEATURES             Location/Qualifiers
     source          1..2967
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     743..760
                     /label=L4440
                     /note="L4440 vector, forward primer"
     CDS             847..1554
                     /codon_start=1
                     /product="reverse tetracycline transactivator 3"
                     /label=rtTA3
                     /note="activates expression from Tet-responsive promoters 
                     in the presence of doxycycline"
                     /translation="MSRLDKSKVINGALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALPIEMLDRHHTHFCPLEGESWQDFLRNNAKSYRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPTDALDDFDLDMLPA
                     DALDDFDLDMLPG"
     primer_bind     complement(1594..1612)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1680..1784
                     /gene="bla"
                     /label=AmpR promoter
     CDS             2006..2797
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
      781 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tcactctgtg gtctcaaggt
      841 gccaccatgt ctaggctgga caagagcaaa gtcataaacg gagctctgga attactcaat
      901 ggtgtcggta tcgaaggcct gacgacaagg aaactcgctc aaaagctggg agttgagcag
      961 cctaccctgt actggcacgt gaagaacaag cgggccctgc tcgatgccct gccaatcgag
     1021 atgctggaca ggcatcatac ccacttctgc cccctggaag gcgagtcatg gcaagacttt
     1081 ctgcggaaca acgccaagtc ataccgctgt gctctcctct cacatcgcga cggggctaaa
     1141 gtgcatctcg gcacccgccc aacagagaaa cagtacgaaa ccctggaaaa tcagctcgcg
     1201 ttcctgtgtc agcaaggctt ctccctggag aacgcactgt acgctctgtc cgccgtgggc
     1261 cactttacac tgggctgcgt attggaggaa caggagcatc aagtagcaaa agaggaaaga
     1321 gagacaccta ccaccgattc tatgccccca cttctgagac aagcaattga gctgttcgac
     1381 cggcagggag ccgaacctgc cttccttttc ggcctggaac taatcatatg tggcctggag
     1441 aaacagctaa agtgcgaaag cggcgggccg accgacgccc ttgacgattt tgacttagac
     1501 atgctcccag ccgatgccct tgacgatttt gaccttgaca tgctccccgg gtaagctttg
     1561 agaccacgaa gtggctcttc agtggacgaa agggcctcgt gatacgccta tttttatagg
     1621 ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc
     1681 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     1741 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgcgc tcacgcaact
     1801 ggtccagaac cttgaccgaa cgcagcggtg gtaacggcgc agtggcggtt ttcatggctt
     1861 gttatgactg tttttttttg gggtacagtc tatgcctcgg gcatccaagc agcaagcgcg
     1921 ttacgccgtg ggtcgatgtt tgatgttatg gagcagcaac gatgttacgc agcagggcag
     1981 tcgccctaaa acaaagttaa acatcatgag ggaagcggtg atcgccgaag tatcgactca
     2041 actatcagag gtagttggcg tcatcgagcg ccatctcgaa ccgacgttgc tggccgtaca
     2101 tttgtacggc tccgcagtgg atggcggcct gaagccacac agcgatattg atttgctggt
     2161 tacggtgacc gtaaggcttg atgaaacaac gcggcgagct ttgatcaacg accttttgga
     2221 aacttcggct tcccctggag agagcgagat tctccgcgct gtagaagtca ccattgttgt
     2281 gcacgacgac atcattccgt ggcgttatcc agctaagcgc gaactgcaat ttggagaatg
     2341 gcagcgcaat gacattcttg caggtatctt cgagccagcc acgatcgaca ttgatctggc
     2401 tatcttgctg acaaaagcaa gagaacatag cgttgccttg gtaggtccag cggcggagga
     2461 actctttgat ccggttcctg aacaggatct atttgaggcg ctaaatgaaa ccttaacgct
     2521 atggaactcg ccgcccgact gggctggcga tgagcgaaat gtagtgctta cgttgtcccg
     2581 catttggtac agcgcagtaa ccggcaaaat cgcgccgaag gatgtcgctg ccgactgggc
     2641 aatggagcgc ctgccggccc agtatcagcc cgtcatactt gaagctagac aggcttatct
     2701 tggacaagaa gaagatcgct tggcctcgcg cgcagatcag ttggaagaat ttgtccatta
     2761 cgtaaaaggc gagatcacca aggtagtcgg caaataactg tcagaccaag tttactcata
     2821 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     2881 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     2941 ccccgtagaa aagatcaaag gatcttc
//
