LOCUS       62056_18310        5113 bp DNA     circular SYN 16-AUG-2021
DEFINITION  Cloning vector pOtta_sfGFP__pOsym_mKate2, complete sequence.
ACCESSION   MZ198787
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5113)
  AUTHORS   Groseclose TM, Hersey AN, Huang BD, Wilson CJ.
  TITLE     Direct Submission
  JOURNAL   
REFERENCE   2  (bases 1 to 5113)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Submitted 
            (13-MAY-2021) Chemical "
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..5113
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(400..494)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(528..1319)
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
     regulatory      1506..1534
                     /label=trc promoter
                     /note="trc promoter"
                     /regulatory_class="promoter"
     regulatory      1506..1511
                     /label=trc promoter -35 hexamer
                     /note="trc promoter -35 hexamer"
                     /regulatory_class="promoter"
     regulatory      1512..1528
                     /label=operator Otta
                     /note="operator Otta"
                     /regulatory_class="other"
     regulatory      1529..1534
                     /label=trc promoter -10 hexamer
                     /note="trc promoter -10 hexamer"
                     /regulatory_class="promoter"
     regulatory      1553..1564
                     /label=strong bacterial RBS
                     /note="strong bacterial RBS"
                     /regulatory_class="ribosome_binding_site"
     CDS             1571..2284
                     /label=superfolder GFP
                     /note="GFP variant that folds robustly even when fused to
                     poorly folded proteins (Nager et al., 2011)"
     terminator      2299..2385
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     CDS             complement(2926..3873)
                     /label=Rep101
                     /note="RepA protein needed for replication with the pSC101 
                     origin"
     rep_origin      complement(3921..4143)
                     /direction=LEFT
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     protein_bind    4256..4275
                     /label=lac operator (symmetric)
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG). The 
                     symmetric lac operator was optimized for tight binding of 
                     lac repressor."
     regulatory      4275..4280
                     /label=trc promoter -10 hexamer
                     /note="trc promoter -10 hexamer"
                     /regulatory_class="promoter"
     regulatory      4299..4310
                     /label=strong bacterial RBS
                     /note="strong bacterial RBS"
                     /regulatory_class="ribosome_binding_site"
     CDS             4317..5012
                     /label=mKate2
                     /note="monomeric far-red fluorescent protein (Shcherbo et
                     al., 2009)"
     terminator      complement(5030..5073)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
ORIGIN
        1 gtagcaccct cgcaagctcg ggcaaatcgc tgaatattcc ttttgtctcc gaccatcagg
       61 cacctgagtc gctgtctttt tcgtgacatt cagttcgctg cgctcacggc tctggcagtg
      121 aatgggggta aatggcacta caggcgcctt ttatggattc atgcaaggaa actacccata
      181 atacaagaaa agcccgtcac gggcttctca gggcgtttta tggcgggtct gctatgtggt
      241 gctatctgac tttttgctgt tcagcagttc ctgccctctg attttccagt ctgaccactt
      301 cggattatcc cgtgacaggt cattcagact ggctaatgca cccagtaagg cagcggtatc
      361 atcaacaggc ttacccgtct tactgtccct agtgcttgga ttctcaccaa taaaaaacgc
      421 ccggcggcaa ccgagcgttc tgaacaaatc cagatggagt tctgaggtca ttactggatc
      481 tatcaacagg agtccaagcg agctctcgaa ccccagagtc ccgctcagaa gaactcgtca
      541 agaaggcgat agaaggcgat gcgctgcgaa tcgggagcgg cgataccgta aagcacgagg
      601 aagcggtcag cccattcgcc gccaagctct tcagcaatat cacgggtagc caacgctatg
      661 tcctgatagc ggtccgccac acccagccgg ccacagtcga tgaatccaga aaagcggcca
      721 ttttccacca tgatattcgg caagcaggca tcgccatggg tcacgacgag atcctcgccg
      781 tcgggcatgc gcgccttgag cctggcgaac agttcggctg gcgcgagccc ctgatgctct
      841 tcgtccagat catcctgatc gacaagaccg gcttccatcc gagtacgtgc tcgctcgatg
      901 cgatgtttcg cttggtggtc gaatgggcag gtagccggat caagcgtatg cagccgccgc
      961 attgcatcag ccatgatgga tactttctcg gcaggagcaa ggtgagatga caggagatcc
     1021 tgccccggca cttcgcccaa tagcagccag tcccttcccg cttcagtgac aacgtcgagc
     1081 acagctgcgc aaggaacgcc cgtcgtggcc agccacgata gccgcgctgc ctcgtcctgc
     1141 agttcattca gggcaccgga caggtcggtc ttgacaaaaa gaaccgggcg cccctgcgct
     1201 gacagccgga acacggcggc atcagagcag ccgattgtct gttgtgccca gtcatagccg
     1261 aatagcctct ccacccaagc ggccggagaa cctgcgtgca atccatcttg ttcaatcatg
     1321 cgaaacgatc ctcatcctgt ctcttgatca gatcttgatc ccctgcgcca tcagatcctt
     1381 ggcggcaaga aagccatcca gtttactttg cagggcttcc caaccttacc agagggcgcc
     1441 ccagctggca attccgacgt ccaacatatg gcaccggagg ctttcgtctt cacctcgaga
     1501 attgtttgac attttaagcg cttaaaatta taatctgagc acagaattca ttaaagagga
     1561 gaaaggtacc atgagcaaag gagaagaact tttcactgga gttgtcccaa ttcttgttga
     1621 attagatggt gatgttaatg ggcacaaatt ttctgtccgt ggagagggtg aaggtgatgc
     1681 tacaaacgga aagcttaccc ttaaatttat ttgcactact ggaaaactac ctgttccatg
     1741 gccaacactt gtcactactc tcacttatgg tgttcaatgc ttttcccgtt atccggatca
     1801 tatgaaacgg catgactttt tcaagagtgc catgcccgaa ggttatgtac aggaacgcac
     1861 tatatctttc aaagatgacg ggacttacaa gacgcgtgct gaagtcaagt ttgaaggtga
     1921 tacccttgtt aatcgtatcg agttaaaagg tattgatttt aaagaagatg gaaacattct
     1981 cggacacaaa cttgagtaca actttaactc acacaatgta tacatcacgg cagacaaaca
     2041 aaagaatgga atcaaagcta acttcaaaat tcgccacaac gttgaagatg gatccgttca
     2101 actagcagac cattatcaac aaaatactcc aattggcgat ggccctgtcc ttttaccaga
     2161 caaccattac ctgtcgacac aatctgtcct ttcgaaagat cccaacgaaa agcgtgacca
     2221 catggtcctt cttgagtttg taactgctgc tgggattaca catggcatgg atgagctcta
     2281 caaataatct agaggcatca aataaaacga aaggctcagt cgaaagactg ggcctttcgt
     2341 tttatctgtt gtttgtcggt gaacgctctc ctgagtagga caaatccgcc gccctagacc
     2401 tagggtacgg gttttgctgc ccgcaaacgg gctgttctgg tgttgctagt ttgttatcag
     2461 aatcgcagat ccggcttcag ccggtttgcc ggctgaaagc gctatttctt ccagaattgc
     2521 catgattttt tccccacggg aggcgtcact ggctcccgtg ttgtcggcag ctttgattcg
     2581 ataagcagca tcgcctgttt caggctgtct atgtgtgact gttgagctgt aacaagttgt
     2641 ctcaggtgtt caatttcatg ttctagttgc tttgttttac tggtttcacc tgttctatta
     2701 ggtgttacat gctgttcatc tgttacattg tcgatctgtt catggtgaac agctttaaat
     2761 gcaccaaaaa ctcgtaaaag ctctgatgta tctatctttt ttacaccgtt ttcatctgtg
     2821 catatggaca gttttccctt tgatatctaa cggtgaacag ttgttctact tttgtttgtt
     2881 agtcttgatg cttcactgat agatacaaga gccataagaa cctcagatcc ttccgtattt
     2941 agccagtatg ttctctagtg tggttcgttg tttttgcgtg agccatgaga acgaaccatt
     3001 gagatcatgc ttactttgca tgtcactcaa aaattttgcc tcaaaactgg tgagctgaat
     3061 ttttgcagtt aaagcatcgt gtagtgtttt tcttagtccg ttacgtaggt aggaatctga
     3121 tgtaatggtt gttggtattt tgtcaccatt catttttatc tggttgttct caagttcggt
     3181 tacgagatcc atttgtctat ctagttcaac ttggaaaatc aacgtatcag tcgggcggcc
     3241 tcgcttatca accaccaatt tcatattgct gtaagtgttt aaatctttac ttattggttt
     3301 caaaacccat tggttaagcc ttttaaactc atggtagtta ttttcaagca ttaacatgaa
     3361 cttaaattca tcaaggctaa tctctatatt tgccttgtga gttttctttt gtgttagttc
     3421 ttttaataac cactcataaa tcctcataga gtatttgttt tcaaaagact taacatgttc
     3481 cagattatat tttatgaatt tttttaactg gaaaagataa ggcaatatct cttcactaaa
     3541 aactaattct aatttttcgc ttgagaactt ggcatagttt gtccactgga aaatctcaaa
     3601 gcctttaacc aaaggattcc tgatttccac agttctcgtc atcagctctc tggttgcttt
     3661 agctaataca ccataagcat tttccctact gatgttcatc atctgagcgt attggttata
     3721 agtgaacgat accgtccgtt ctttccttgt agggttttca atcgtggggt tgagtagtgc
     3781 cacacagcat aaaattagct tggtttcatg ctccgttaag tcatagcgac taatcgctag
     3841 ttcatttgct ttgaaaacaa ctaattcaga catacatctc aattggtcta ggtgatttta
     3901 atcactatac caattgagat gggctagtca atgataatta ctagtccttt tcccgggaga
     3961 tctgggtatc tgtaaattct gctagacctt tgctggaaaa cttgtaaatt ctgctagacc
     4021 ctctgtaaat tccgctagac ctttgtgtgt tttttttgtt tatattcaag tggttataat
     4081 ttatagaata aagaaagaat aaaaaaagat aaaaagaata gatcccagcc ctgtgtataa
     4141 ctcactactt tagtcagttc cgcagtatta caaaaggatg tcgcaaacgc tgtttgctcc
     4201 tctacaaaac agaccttaaa accctaaagg cgtcttcacc tcgagaattg tttgacattg
     4261 tgagcgctca caattataat ctgagcacag aattcattaa agaggagaaa ggtaccatgg
     4321 tatctgaact gatcaaggaa aacatgcata tgaagttgta tatggaaggt accgtgaaca
     4381 accatcattt caagtgtact agtgaaggtg aaggaaaacc gtatgaagga actcagacaa
     4441 tgcgtattaa agctgtagaa ggtggaccgc tgccttttgc tttcgatatc ttggcaacct
     4501 ctttcatgta tggtagtaag actttcatca accatacaca gggaatcccg gatttcttta
     4561 aacaatcgtt tcctgaaggt ttcacatggg aacgtgtaac cacttatgaa gatggaggag
     4621 tactgaccgc cactcaggat accagcctgc aagatggatg tttgatctat aacgtaaaga
     4681 tccgtggtgt gaacttcccg tcgaatggac ctgtaatgca gaagaaaacc ttgggttggg
     4741 aagccagcac agaaaccctg tatccggctg atggaggact ggaaggacgt gcagatatgg
     4801 ccctgaaact ggtaggtgga ggacatctga tctgtaactt gaagacaacc tatcgttcta
     4861 aaaaaccggc aaagaacctg aagatgcctg gtgtatatta tgtggatcgt cgtttggaac
     4921 gtatcaaaga agccgataaa gaaacatatg tggaacaaca tgaagtagcc gtggctcgtt
     4981 attgtgatct gccttctaaa ttgggacatc gttaatctag aggcatcaaa taaaacgaaa
     5041 ggctcagtcg aaagactggg cctttcgttt tatctgttgt ttgtcggtga acgctctcct
     5101 gagtaggaca aat
//
