LOCUS       Exported                4242 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Expresses mCherry under the PLtetO-1 promoter.
ACCESSION   .
VERSION     .
KEYWORDS    pBL6
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4242)
  AUTHORS   Olson EJ, Hartsough LA, Landry BP, Shroff R, Tabor JJ
  TITLE     Characterizing bacterial gene circuit dynamics with optically 
            programmed gene expression signals.
  JOURNAL   Nat Methods. 2014 Mar 9. doi: 10.1038/nmeth.2884.
  PUBMED    24608181
REFERENCE   2  (bases 1 to 4242)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..4242
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     complement(72..89)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     CDS             complement(351..1301)
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLRNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKHDLNGSFSWLTQKQRTTLENILAKYGRI"
     rep_origin      1349..1571
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     terminator      2063..2157
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(2187..3047)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     2810..2829
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3048..3152)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3217..3235
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      3239..3282
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     terminator      3303..3330
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      3346..3417
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(3426..4136)
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(3704..3723)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     3973..3991
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     RBS             4144..4155
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     promoter        complement(4165..4238)
                     /label=PLtetO-1 promoter
                     /note="modified phage lambda PL promoter with tet operator 
                     sites (Lutz and Bujard, 1997)"
     protein_bind    4195..4213
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    4220..4238
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
ORIGIN
        1 aagggcaagg tgtcaccacc ctgccctttt tctttaaaac cgaaaagatt acttcgcgtt
       61 atgcaggctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
      121 tcagctcact caaaggcggt aatctcgagg ttacattgtc gatctgttca tggtgaacag
      181 ctttaaatgc accaaaaact cgtaaaagct ctgatgtatc tatctttttt acaccgtttt
      241 catctgtgca tatggacagt tttccctttg atatctaacg gtgaacagtt gttctacttt
      301 tgtttgttag tcttgatgct tcactgatag atacaagagc cataagaacc tcagatcctt
      361 ccgtatttag ccagtatgtt ctctagtgtg gttcgttgtt tttgcgtgag ccatgagaac
      421 gaaccattga gatcatgctt actttgcatg tcactcaaaa attttgcctc aaaactggtg
      481 agctgaattt ttgcagttaa agcatcgtgt agtgtttttc ttagtccgtt acgtaggtag
      541 gaatctgatg taatggttgt tggtattttg tcaccattca tttttatctg gttgttctca
      601 agttcggtta cgagatccat ttgtctatct agttcaactt ggaaaatcaa cgtatcagtc
      661 gggcggcctc gcttatcaac caccaatttc atattgctgt aagtgtttaa atctttactt
      721 attggtttca aaacccattg gttaagcctt ttaaactcat ggtagttatt ttcaagcatt
      781 aacatgaact taaattcatc aaggctaatc tctatatttg ccttgtgagt tttcttttgt
      841 gttagttctt ttaataacca ctcataaatc ctcatagagt atttgttttc aaaagactta
      901 acatgttcca gattatattt tatgaatttt tttaactgga aaagataagg caatatctct
      961 tcactaaaaa ctaattctaa tttttcgctt gagaacttgg catagtttgt ccactggaaa
     1021 atctcaaagc ctttaaccaa aggattcctg atttccacag ttctcgtcat cagctctctg
     1081 gttgctttag ctaatacacc ataagcattt tccctactga tgttcatcat ctgagcgtat
     1141 tggttataag tgaacgatac cgtccgttct ttccttgtag ggttttcaat cgtggggttg
     1201 agtagtgcca cacagcataa aattagcttg gtttcatgct ccgttaagtc atagcgacta
     1261 atcgctagtt catttgcttt gaaaacaact aattcagaca tacatctcaa ttggtctagg
     1321 tgattttaat cactatacca attgagatgg gctagtcaat gataattaca tgtccttttc
     1381 ctttgagttg tgggtatctg taaattctgc tagacctttg ctggaaaact tgtaaattct
     1441 gctagaccct ctgtaaattc cgctagacct ttgtgtgttt tttttgttta tattcaagtg
     1501 gttataattt atagaataaa gaaagaataa aaaaagataa aaagaataga tcccagccct
     1561 gtgtataact cactacttta gtcagttccg cagtattaca aaaggatgtc gcaaacgctg
     1621 tttgctcctc tacaaaacag accttaaaac cctaaaggct taagtagcac cctcgcaagc
     1681 tcgggcaaat cgctgaatat tccttttgtc tccgaccatc aggcacctga gtcgctgtct
     1741 ttttcgtgac attcagttcg ctgcgctcac ggctctggca gtgaatgggg gtaaatggca
     1801 ctacaggcgc cttttatgga ttcatgcaag gaaactaccc ataatacaag aaaagcccgt
     1861 cacgggcttc tcagggcgtt ttatggcggg tctgctatgt ggtgctatct gactttttgc
     1921 tgttcagcag ttcctgccct ctgattttcc agtctgacca cttcggatta tcccgtgaca
     1981 ggtcattcag actggctaat gcacccagta aggcagcggt atcatcaaca ggcttacccg
     2041 tcttactgtc cctagtgctt ggattctcac caataaaaaa cgcccggcgg caaccgagcg
     2101 ttctgaacaa atccagatgg agttctgagg tcattactgg atctatcaac aggagtccaa
     2161 gcgagctcgt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     2221 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     2281 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     2341 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     2401 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     2461 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     2521 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     2581 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     2641 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     2701 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     2761 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     2821 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     2881 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     2941 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     3001 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     3061 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     3121 atttagaaaa ataaacaaat aggggttccg cgcacatttc ccatggtgcc acctgacgtc
     3181 taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcagaat
     3241 ttcagataaa aaaaatcctt agctttcgct aaggatgatt tctggaattc gcagagtata
     3301 aacgcagaaa ggcccacccg aaggtgagcc agtgtgactc tagtagagag cgttcaccga
     3361 caaacaacag ataaaacgaa aggcccagtc tttcgactga gcctttcgtt ttatttgatg
     3421 cctggttact tgtacagctc gtccatgccg ccggtggagt ggcggccctc ggcgcgttcg
     3481 tactgttcca cgatggtgta gtcctcgttg tgggaggtga tgtccaactt gatgttgacg
     3541 ttgtaggcgc cgggcagctg cacgggcttc ttggccttgt aggtggtctt gacctcagcg
     3601 tcgtagtggc cgccgtcctt cagcttcagc ctctgcttga tctcgccctt cagggcgccg
     3661 tcctcggggt acatccgctc ggaggaggcc tcccagccca tggtcttctt ctgcattacg
     3721 gggccgtcgg aggggaagtt ggtgccgcgc agcttcacct tgtagatgaa ctcgccgtcc
     3781 tgcagggagg agtcctgggt cacggtcacc acgccgccgt cctcgaagtt catcacgcgc
     3841 tcccacttga agccctcggg gaaggacagc ttcaagtagt cggggatgtc ggcggggtgc
     3901 ttcacgtagg ccttggagcc gtacatgaac tgaggggaca ggatgtccca ggcgaagggc
     3961 agggggccac ccttggtcac cttcagcttg gcggtctggg tgccctcgta ggggcggccc
     4021 tcgccctcgc cctcgatctc gaactcgtgg ccgttcacgg agccctccat gtgcaccttg
     4081 aagcgcatga actccttgat gatggccatg ttatcctcct cgcccttgct caccatgggt
     4141 acctttctcc tctttaatga attcggtcag tgcgtcctgc tgatgtgctc agtatctcta
     4201 tcactgatag ggatgtcaat ctctatcact gatagggata ct
//
