LOCUS       62056_14140        4770 bp DNA     circular SYN 27-JUL-2020
DEFINITION  Cloning vector pLH401, complete sequence.
ACCESSION   MN617156
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4770)
  AUTHORS   Hartsough LA, Kotlajich MV, Han B, Lin C-C.J., Gambill L, Wang MC, 
            Tabor JJ.
  TITLE     Optogenetic control of gut bacterial metabolism
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4770)
  AUTHORS   Hartsough LA, Kotlajich MV, Han B, Lin C-C.J., Gambill L, Wang MC, 
            Tabor JJ.
  TITLE     Direct Submission
  JOURNAL   Submitted (26-OCT-2019) Bioengineering, Rice University, 6500 Main 
            St., Houston, TX 77030, USA
REFERENCE   3  (bases 1 to 4770)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (26-OCT-2019) Bioengineering, Rice University, 6500 Main St., 
            Houston, TX 77030, USA"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..4770
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(72..660)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     terminator      complement(748..842)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(866..1522)
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
     promoter        complement(1523..1625)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding
                     chloramphenicol acetyltransferase"
     regulatory      1651..1685
                     /label=constitutive promoter J23114
                     /note="constitutive promoter J23114"
                     /regulatory_class="promoter"
     regulatory      1693..1706
                     /label=RBS pprotet
                     /note="RBS pprotet"
                     /regulatory_class="ribosome_binding_site"
     misc_feature    1695..1706
                     /label=B0034
                     /note="B0034"
     CDS             1714..2421
                     /label=mCherry
                     /note="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
     misc_feature    2425..2484
                     /label=L3S1P52
                     /note="L3S1P52"
     regulatory      2564..2598
                     /label=constitutive promoter J23100
                     /note="constitutive promoter J23100"
                     /regulatory_class="promoter"
     regulatory      2599..2632
                     /label=sRBS
                     /note="sRBS"
                     /regulatory_class="ribosome_binding_site"
     CDS             2633..3337
                     /codon_start=1
                     /transl_table=11
                     /product="CcaR"
                     /label=CcaR
                     /protein_id="QLL27763.1"
                     /translation="MRILLVEDDLPLAETLAEALSDQLYTVDIATDASLAWDYASRLEY
                     DLVILDVMLPELDGITLCQKWRSHSYLMPILMMTARDTINDKITGLDAGADDYVVKPVD
                     LGELFARVRALLRRGCATCQPVLEWGPIRLDPSTYEVSYDNEVLSLTRKEYSILELLLR
                     NGRRVLSRSMIIDSIWKLESPPEEDTVKVHVRSLRQKLKSAGLSADAIETVHGIGYRLA
                     NLTEKSLCQGKN"
     terminator      3358..3429
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      3445..3472
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     misc_feature    complement(3584..3595)
                     /label=stop region
                     /note="stop region"
     regulatory      3596..3767
                     /label=truncated PcpcG2
                     /note="truncated PcpcG2"
                     /regulatory_class="promoter"
     regulatory      3768..3785
                     /label=B0034
                     /note="B0034"
                     /regulatory_class="ribosome_binding_site"
     CDS             3786..4499
                     /label=superfolder GFP
                     /note="GFP variant that folds robustly even when fused to
                     poorly folded proteins (Pedelacq et al., 2006)"
     terminator      complement(4529..4572)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
ORIGIN
        1 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
       61 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
      121 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
      181 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
      241 ccttcgggaa gcgtggcgct ttctcaatgc tcacgctgta ggtatctcag ttcggtgtag
      301 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
      361 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
      421 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
      481 aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg
      541 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
      601 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
      661 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
      721 gggattttgg tcatgactag tgcttggatt ctcaccaata aaaaacgccc ggcggcaacc
      781 gagcgttctg aacaaatcca gatggagttc tgaggtcatt actggatcta tcaacaggag
      841 tccaagcgag ctcgatatca aattacgccc cgccctgcca ctcatcgcag tactgttgta
      901 attcattaag cattctgccg acatggaagc catcacagac ggcatgatga acctgaatcg
      961 ccagcggcat cagcaccttg tcgccttgcg tataatattt gcccatggtg aaaacggggg
     1021 cgaagaagtt gtccatattg gccacgttta aatcaaaact ggtgaaactc acccagggat
     1081 tggctgagac gaaaaacata ttctcaataa accctttagg gaaataggcc aggttttcac
     1141 cgtaacacgc cacatcttgc gaatatatgt gtagaaactg ccggaaatcg tcgtggtatt
     1201 cactccagag cgatgaaaac gtttcagttt gctcatggaa aacggtgtaa caagggtgaa
     1261 cactatccca tatcaccagc tcaccgtctt tcattgccat acgaaattcc ggatgagcat
     1321 tcatcaggcg ggcaagaatg tgaataaagg ccggataaaa cttgtgctta tttttcttta
     1381 cggtctttaa aaaggccgta atatccagct gaacggtctg gttataggta cattgagcaa
     1441 ctgactgaaa tgcctcaaaa tgttctttac gatgccattg ggatatatca acggtggtat
     1501 atccagtgat ttttttctcc attttagctt ccttagctcc tgaaaatctc gataactcaa
     1561 aaaatacgcc cggtagtgat cttatttcat tatggtgaaa gttggaacct cttacgtgcc
     1621 gatcaacgtc tcattttcgc cagatatcga tttatggcta gctcagtcct aggtacaatg
     1681 ctagcgaatt cattaaagag gagaaaggta cccatggtga gcaagggcga ggaggataac
     1741 atggccatca tcaaggagtt catgcgcttc aaggtgcaca tggagggctc cgtgaacggc
     1801 cacgagttcg agatcgaggg cgagggcgag ggccgcccct acgagggcac ccagaccgcc
     1861 aagctgaagg tgaccaaggg tggccccctg cccttcgcct gggacatcct gtcccctcag
     1921 ttcatgtacg gctccaaggc ctacgtgaag caccccgccg acatccccga ctacttgaag
     1981 ctgtccttcc ccgagggctt caagtgggag cgcgtgatga acttcgagga cggcggcgtg
     2041 gtgaccgtga cccaggactc ctccctgcag gacggcgagt tcatctacaa ggtgaagctg
     2101 cgcggcacca acttcccctc cgacggcccc gtaatgcaga agaagaccat gggctgggag
     2161 gcctcctccg agcggatgta ccccgaggac ggcgccctga agggcgagat caagcagagg
     2221 ctgaagctga aggacggcgg ccactacgac gctgaggtca agaccaccta caaggccaag
     2281 aagcccgtgc agctgcccgg cgcctacaac gtcaacatca agttggacat cacctcccac
     2341 aacgaggact acaccatcgt ggaacagtac gaacgcgccg agggccgcca ctccaccggc
     2401 ggcatggacg agctgtacaa gtaagctttc taactaaaaa ggcctcccaa atcggggggc
     2461 cttttttatt gataacaaaa cgctcgtcta agaaaccatt attatcatga cattaaccta
     2521 taaaaatagg cgtatcacga ggccctttcg tcttcacctc gagttgacgg ctagctcagt
     2581 cctaggtaca gtgctagctg ttaaacgctc ctcgtaagaa cacgggaaag caatgagaat
     2641 tcttttagtg gaggatgatt tgccgctggc ggaaaccctt gctgaagcat tgagtgacca
     2701 gctttacacc gttgatattg ccaccgacgc ttccctcgcc tgggactatg cctcccgact
     2761 ggaatatgac ctcgttattt tggatgtgat gctgccggag ttggacggga ttaccctctg
     2821 tcaaaaatgg cgatcgcaca gttatttaat gccaattttg atgatgacag ccagggatac
     2881 gatcaatgat aaaatcacgg gcttggatgc gggggcggat gattatgtgg tcaagccagt
     2941 ggatttgggg gagttatttg ccagggtgcg agctttgttg cgtcggggtt gtgcaacgtg
     3001 ccaaccagtt ttagagtggg ggccaatcag gttggatcca agcacctatg aagttagtta
     3061 tgacaatgag gttttgtctt tgacccgcaa ggaatacagc attctggaat tactactccg
     3121 caatggccgt cgggtgctaa gtcggagcat gattatcgat agtatctgga agttggagag
     3181 tcccccagag gaagatacgg ttaaggtgca tgtgcggagt ttgcgacaaa aattaaaaag
     3241 tgccggttta tcagcagatg ccattgaaac ggtccatggc attgggtatc gtctggccaa
     3301 tttaacggaa aaatctttgt gccaagggaa aaactagtaa taatctagac caggcatcaa
     3361 ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt ttatctgttg tttgtcggtg
     3421 aacgctctct actagagtca cactggctca ccttcgggtg ggcctttctg cgtttatagt
     3481 aaatcactgc ataattcgtg tcgctcaagg cgcactcccg ttctggataa tgttttttgc
     3541 gccgacatca taacggttct ggcaaatatt ctgaaatgag ctggttagct agtcaagccc
     3601 attgtgcttt tctctatcaa cctcagctta cctgaagggg tgaacaggtc tgggttaatt
     3661 catgttgcga aatgtaacag ttttagtcgc atcagctaac tttccgattt ctttacgatt
     3721 ttctccccct tttcttcaat tttactttgt taggatcgca tttttaaaaa gaggagaaat
     3781 actagatgcg taaaggcgaa gagctgttca ctggtgtcgt ccctattctg gtggaactgg
     3841 atggtgatgt caacggtcat aagttttccg tgcgtggcga gggtgaaggt gacgcaacta
     3901 atggtaaact gacgctgaag ttcatctgta ctactggtaa actgccggta ccttggccga
     3961 ctctggtaac gacgctgact tatggtgttc agtgctttgc tcgttatccg gaccatatga
     4021 agcagcatga cttcttcaag tccgccatgc cggaaggcta tgtgcaggaa cgcacgattt
     4081 cctttaagga tgacggcacg tacaaaacgc gtgcggaagt gaaatttgaa ggcgataccc
     4141 tggtaaaccg cattgagctg aaaggcattg actttaaaga agacggcaat atcctgggcc
     4201 ataagctgga atacaatttt aacagccaca atgtttacat caccgccgat aaacaaaaaa
     4261 atggcattaa agcgaatttt aaaattcgcc acaacgtgga ggatggcagc gtgcagctgg
     4321 ctgatcacta ccagcaaaac actccaatcg gtgatggtcc tgttctgctg ccagacaatc
     4381 actatctgag cacgcaaagc gttctgtcta aagatccgaa cgagaaacgc gatcatatgg
     4441 ttctgctgga gttcgtaacc gcagcgggca tcacgcatgg tatggatgaa ctgtacaaat
     4501 gatgataata atctagacca ggcatcaaat aaaacgaaag gctcagtcga aagactgggc
     4561 ctttcgtttt atctgttgtt tgtcggtgaa cgctctctac tagagtcaca ctggctcacc
     4621 ttcgggtggg cctttctgcg tttataggta cccgtggatc ctctagagtc gacctgcagg
     4681 catgcaagct tcctaggcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     4741 atacggttat ccacagaatc aggggataac
//
