LOCUS       62056_20180        4543 bp DNA     circular SYN 27-JUL-2020
DEFINITION  Cloning vector pSR49-2, complete sequence.
ACCESSION   MN617164
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4543)
  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  JOURNAL   Unpublished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 4543)
  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 4543)
  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            "Unpublished"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..4543
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      1..148
                     /note="specR promoter; from KD13"
                     /regulatory_class="promoter"
     CDS             149..937
                     /label=SmR
                     /note="aminoglycoside adenylyltransferase (Murphy, 1985)"
     terminator      952..1046
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     rep_origin      1160..1705
                     /direction=RIGHT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin."
     primer_bind     1835..1855
                     /label=sequencing primer fwSR26
                     /note="sequencing primer fwSR26"
     misc_feature    1961..1995
                     /label=constitutive promoter J23106
                     /note="constitutive promoter J23106"
     misc_feature    1996..2030
                     /label=B0034
                     /note="B0034"
     CDS             2031..4292
                     /codon_start=1
                     /transl_table=11
                     /product="CcaS"
                     /label=CcaS
                     /protein_id="QLL27778.1"
                     /translation="MGKFLIPIEFVFLAIAMTCYLWHRQNQERRRIEISIKQQTQRERF
                     INQITQHIRQSLNLETVLNTTVAEVKTLLQVDRVLIYRIWQDGTGSAITESVNANYPSI
                     LGRTFSDEVFPVEYHQAYTKGKVRAINDIDQDDIEICLADFVKQFGVKSKLVVPILQHN
                     RASSLDNESEFPYLWGLLITHQCAFTRPWQPWEVELMKQLANQVAIAIQQSELYEQLQQ
                     LNKDLENRVEKRTQQLAATNQSLRMEISERQKTEAALRHTNHTLQSLIAASPRGIFTLN
                     LADQIQIWNPTAERIFGWTETEIIAHPELLTSNILLEDYQQFKQKVLSGMVSPSLELKC
                     QKKDGSWIEIVLSAAPLLDSEENIAGLVAVVADITEQKRQAEQIRLLQSVVVNTNDAVV
                     ITEAEPIDDPGPRILYVNEAFTKITGYTAEEMLGKTPRVLQGPKTSRTELDRVRQAISQ
                     WQSVTVEVINYRKDGSEFWVEFSLVPVANKTGFYTHWIAVQRDVTERRRTEEVRLALER
                     EKELSRLKTRFFSMASHEFRTPLSTALAAAQLLENSEVAWLDPDKRSRNLHRIQNSVKN
                     MVQLLDDILIINRAEAGKLEFNPNWLDLKLLFQQFIEEIQLSVSDQYYFDFICSAQDTK
                     ALVDERLVRSILSNLLSNAIKYSPGGGQIKIALSLDSEQIIFEVTDQGIGISPEDQKQI
                     FEPFHRGKNVRNITGTGLGLMVAKKCVDLHSGSILLKSAVDQGTTVTICLKRYNHLPRA
                     "
     misc_feature    2187..2741
                     /note="GAF domain; derived from Synechocystis PCC6803 CcaS"
     terminator      4313..4384
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      4400..4427
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
ORIGIN
        1 agcaagcgaa ccggaattgc cagctggggc gccctctggt aaggttggga agccctgcaa
       61 agtaaactgg atggctttct tgccgccaag gatctgatgg cgcaggggat caagatctga
      121 tcaagagaca ggatgaggat cgtttcgcat gagggaagcg gtgatcgccg aagtatcgac
      181 tcaactatca gaggtagttg gcgtcatcga gcgccatctc gaaccgacgt tgctggccgt
      241 acatttgtac ggctccgcag tggatggcgg cctgaagcca cacagtgata ttgatttgct
      301 ggttacggtg accgtaaggc ttgatgaaac aacgcggcga gctttgatca acgacctttt
      361 ggaaacttcg gcttcccctg gagagagcga gattctccgc gctgtagaag tcaccattgt
      421 tgtgcacgac gacatcattc cgtggcgtta tccagctaag cgcgaactgc aatttggaga
      481 atggcagcgc aatgacattc ttgcaggtat cttcgagcca gccacgatcg acattgatct
      541 ggctatcttg ctgacaaaag caagagaaca tagcgttgcc ttggtaggtc cagcggcgga
      601 ggaactcttt gatccggttc ctgaacagga tctatttgag gcgctaaatg aaaccttaac
      661 gctatggaac tcgccgcccg actgggctgg cgatgagcga aatgtagtgc ttacgttgtc
      721 ccgcatttgg tacagcgcag taaccggcaa aatcgcgccg aaggatgtcg ctgccgactg
      781 ggcaatggag cgcctgccgg cccagtatca gcccgtcata cttgaagcta gacaggctta
      841 tcttggacaa gaagaagatc gcttggcctc gcgcgcagat cagttggaag aatttgtcca
      901 ctacgtgaaa ggcgagatca ccaaggtagt cggcaaataa gagctcgctt ggactcctgt
      961 tgatagatcc agtaatgacc tcagaactcc atctggattt gttcagaacg ctcggttgcc
     1021 gccgggcgtt ttttattggt gagaatccaa gcactagtaa caacttatat cgtatggggc
     1081 tgacttcagg tgctacattt gaagagataa attgcactga aatctagtaa tattttatct
     1141 gattaataag atgatcttct tgagatcgtt ttggtctgcg cgtaatctct tgctctgaaa
     1201 acgaaaaaac cgccttgcag ggcggttttt cgaaggttct ctgagctacc aactctttga
     1261 accgaggtaa ctggcttgga ggagcgcagt caccaaaact tgtcctttca gtttagcctt
     1321 aaccggcgca tgacttcaag actaactcct ctaaatcaat taccagtggc tgctgccagt
     1381 ggtgcttttg catgtctttc cgggttggac tcaagacgat agttaccgga taaggcgcag
     1441 cggtcggact gaacgggggg ttcgtgcata cagtccagct tggagcgaac tgcctacccg
     1501 gaactgagtg tcaggcgtgg aatgagacaa acgcggccat aacagcggaa tgacaccggt
     1561 aaaccgaaag gcaggaacag gagagcgcac gagggagccg ccagggggaa acgcctggta
     1621 tctttatagt cctgtcgggt ttcgccacca ctgatttgag cgtcagattt cgtgatgctt
     1681 gtcagggggg cggagcctat ggaaaaacgg ctttgccgcg gccctctcac ttccctgtta
     1741 agtatcttcc tggcatcttc caggaaatct ccgccccgtt cgtaagccat ttccgctcgc
     1801 cgcagtcgaa cgaccgagcg tagcgagtca gtgagcgagg aagcggaata tatcccctag
     1861 gtggagtctg gcctcaaata caaagtctgc aaagtctcga aatattaaac cagagaataa
     1921 ataattaaac gccaaaatat ttctagcaaa aaccccagtt tttacggcta gctcagtcct
     1981 aggtatagtg ctagctatag aggttaaaat cttaatagga ggaaggcgca atgggcaaat
     2041 ttctaattcc aatcgaattt gtttttctgg cgatcgccat gacctgttat ttatggcaca
     2101 gacaaaacca agaacgccgc aggattgaaa ttagcatcaa gcaacaaacc caacgggaac
     2161 gatttattaa ccaaattacc caacatatcc gccaatcttt aaacttggaa acggttttaa
     2221 ataccaccgt cgctgaagtt aaaaccctgt tgcaagttga tcgagttcta atttatcgca
     2281 tttggcaaga tggcacgggc agcgccatta cggaatcggt gaatgccaat tatcctagta
     2341 ttttagggcg gaccttttcc gatgaagttt ttcccgttga ataccatcaa gcctacacca
     2401 aaggtaaagt acgggccatt aatgacattg accaggatga catagagatt tgcctagctg
     2461 atttcgtcaa acaatttggc gtgaaatcaa aattagtagt gcccattctt caacataatc
     2521 gtgcttcttc cctagataat gaatcagaat ttccctatct ttgggggctg ttaattaccc
     2581 atcaatgtgc ttttacccgg ccatggcaac cgtgggaagt ggagttaatg aaacagctag
     2641 ccaatcaggt cgcgatcgcc atccaacaat cggaattata tgagcaatta cagcaactca
     2701 ataaagattt ggaaaaccga gtcgaaaaac gcacccagca acttgccgcc accaatcaat
     2761 ccctaagaat ggaaatcagt gagcgacaaa aaacggaagc cgctctccgc cacactaacc
     2821 atactctgca atccctgatt gcggcctccc ccaggggtat ttttaccctt aatttagcag
     2881 accaaattca gatttggaat cctacagcag aacgtatttt tggttggaca gaaacagaaa
     2941 ttattgccca tccagaatta ttaacatcca acattttgct ggaagattat cagcaattta
     3001 aacagaaagt tttatcaggc atggtttccc ctagcctaga attaaaatgt caaaaaaaag
     3061 atggtagttg gattgaaatt gtcctttccg ctgctcccct attggatagt gaagaaaata
     3121 ttgccggatt ggtggcggtt gtcgccgata ttaccgagca aaagcggcag gcagaacaaa
     3181 ttcgtttgct acaatccgtt gtggttaata ctaatgatgc ggtggtgatt acggaagcgg
     3241 agcccattga tgatcccggg ccgagaattc tctatgtcaa tgaagcattt actaaaatca
     3301 ccggttatac tgctgaagaa atgctaggca aaaccccccg agttttacag ggaccaaaaa
     3361 ctagtcgcac tgaattagat agggtgcggc aagccattag tcaatggcaa tcagttaccg
     3421 ttgaagtgat taattatcgt aaggatggca gtgagttttg ggtggaattt agtctggtgc
     3481 ccgttgccaa taaaacaggt ttttacaccc attggattgc tgtgcaaagg gatgtcactg
     3541 agcgccgacg cacggaggaa gtccgcctag ctttagaacg ggaaaaagaa ttaagccgcc
     3601 taaaaactcg ttttttctcc atggcttccc atgaatttcg tactcccctc agtacggcct
     3661 tagctgctgc ccaattactg gaaaattctg aagtggcctg gcttgatccc gataagcgta
     3721 gccggaactt acaccgtatt caaaattccg tgaaaaatat ggtacagctc ctggatgata
     3781 ttttaatcat taaccgtgcc gaagcgggca aattggaatt taatcctaat tggttagatt
     3841 tgaaattatt gttccagcaa tttatcgaag aaattcaatt aagtgtcagt gaccaatatt
     3901 attttgactt tatttgtagc gctcaagata cgaaggcatt ggtggatgaa aggttagtgc
     3961 ggtctatttt atctaatctg ttatctaatg cgattaaata ctctcccggg ggagggcaga
     4021 ttaaaattgc cctaagccta gattcggaac agattatttt tgaagtcacc gaccagggca
     4081 ttggcatttc gccagaggac caaaagcaaa tttttgaacc ctttcatcgg ggcaaaaatg
     4141 tcagaaatat tacgggaaca ggactcggtt taatggttgc caagaaatgt gttgacttac
     4201 acagtggcag tatcttgcta aaaagtgcag ttgaccaggg aacaacagtt actatctgtt
     4261 taaaacgcta taaccatttg cctcgagctt agtaataatc tagaccaggc atcaaataaa
     4321 acgaaaggct cagtcgaaag actgggcctt tcgttttatc tgttgtttgt cggtgaacgc
     4381 tctctactag agtcacactg gctcaccttc gggtgggcct ttctgcgttt atagtaaatc
     4441 actgcataat tcgtgtcgat taaaccgcct aagtccccca ggaaaggggg atataacagt
     4501 atagattttg tcagccttca gcttggcttt accgtcaaaa aaa
//
