LOCUS       62056_21595        5191 bp DNA     circular SYN 09-MAY-2021
DEFINITION  Cloning vector pTS9, complete sequence.
ACCESSION   MW835298
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5191)
  AUTHORS   Schnabel T.
  TITLE     Engineering post-translational regulation of glutamine synthetase 
            for controllable ammonia production in the plant-symbiont A. 
            brasilense
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5191)
  AUTHORS   Schnabel T.
  TITLE     Direct Submission
  JOURNAL   Submitted (30-MAR-2021) Bioengineering, Stanford University, 443 Via
            Ortega, Stanford, CA 94305, USA
REFERENCE   3  (bases 1 to 5191)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (30-MAR-2021) Bioengineering, Stanford University, 443 Via Ortega, 
            Stanford, CA 94305, USA"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..5191
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      139..727
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     regulatory      778..812
                     /label=pJ23101
                     /note="pJ23101"
                     /regulatory_class="promoter"
     regulatory      828..842
                     /label=B0030
                     /note="B0030"
                     /regulatory_class="ribosome_binding_site"
     gene            849..1868
                     /gene="pheS*"
                     /label=pheS*
     CDS             849..1868
                     /codon_start=1
                     /transl_table=11
                     /gene="pheS*"
                     /product="phenylalanine--tRNA ligase alpha subunit"
                     /function="toxic in presence of 4-chlorophenylalanine"
                     /label=pheS*
                     /protein_id="QUQ60671.1"
                     /translation="MLDALKDELLSQVNAAGDLAALEEVRVTALGKKGRITGFMKELGG
                     LSPDERRERGQQLNALKDEIAAAIDGRKADLARAHLEARLQAERIDVTLPVRPETEGRI
                     HPISQTIDEMVAIFAEMGFSVAEGPDVEDDFHNFTALNFPPGHPARDMHDTFYLPDAGD
                     KKMLLRTHTSPVQVRTMLNKKPPIRIIAPGRTYRSDYDMTHTPMFHQIEGLVIDEATHM
                     GHLKGCLIEFCRAFFDVDDLPLRFRPSFFPFAEPSAEVDIGCSRKGGELKLGNYGDWLE
                     ILGCGMVHPNVLEACGIDSTKYQGFGFGMGIERVAMLKYGIPDLRTFFEADLRWLKHY"
     regulatory      1885..1997
                     /label=pKan
                     /note="pKan"
                     /regulatory_class="promoter"
     oriT            complement(1885..1936)
                     /direction=LEFT
                     /note="oriT"
     CDS             2004..2795
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
     misc_feature    2830..2873
                     /label=multiple cloning site
                     /note="multiple cloning site"
     CDS             2905..2916
                     /label=Factor Xa site
                     /note="Factor Xa recognition and cleavage site"
     misc_feature    3299..3309
                     /label=threeway stop
                     /note="threeway stop"
     misc_feature    3316..3362
                     /note="insert spacer for knock-out or circuit of interest
                     for knock-in"
     misc_feature    3369..3379
                     /label=threeway stop
                     /note="threeway stop"
     misc_feature    3380..3800
                     /note="downstream overlap to chromosome (here an example 
                     fragment of A. brasilense Sp245)"
     misc_feature    3801..3821
                     /label=multiple cloning site
                     /note="multiple cloning site"
     oriT            complement(3913..4022)
                     /direction=LEFT
                     /label=oriT
                     /note="incP origin of transfer"
     promoter        4194..4298
                     /label=AmpR promoter
     CDS             4299..5156
                     /label=AmpR
                     /note="beta-lactamase"
ORIGIN
        1 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
       61 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
      121 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
      181 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
      241 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
      301 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
      361 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
      421 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
      481 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
      541 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
      601 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
      661 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
      721 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgcg tttaaacttt
      781 acagctagct cagtcctagg tattatgcta gcacgggagc aaccgctatt aaagaggaga
      841 aatactagat gctggacgcc ctgaaagacg aactgctgag ccaagtgaac gcggccggtg
      901 atctggcggc tctagaggaa gtgcgcgtga ccgctctcgg caagaaaggt aggatcaccg
      961 gcttcatgaa agagcttggg ggcttgtcgc cggatgaacg ccgcgaacgt gggcaacagc
     1021 tgaacgcctt gaaggatgaa atcgccgcgg cgatcgacgg gcggaaggcc gacctggccc
     1081 gcgcccatct cgaggcgcgg ttgcaggccg agcgcattga tgtaaccctc ccggtgcgac
     1141 cggagaccga agggcgcatc catcctatct cacaaaccat agacgaaatg gtcgccatct
     1201 tcgctgaaat gggcttttcc gtggcagaag gccccgatgt ggaagacgat tttcataatt
     1261 tcaccgccct taactttcca cccggccacc ccgccaggga catgcatgac accttctatc
     1321 tgcccgacgc aggtgacaag aaaatgctgc tcaggacgca cacctccccg gtgcaggtcc
     1381 gcacgatgct caacaagaag cccccgatcc gcatcattgc tcccggtcgt acgtatcgtt
     1441 cagactatga tatgacccac accccgatgt tccatcaaat cgagggattg gtgatagacg
     1501 aagcaaccca tatgggccat ctgaagggat gtctgatcga attttgtcgc gcgttcttcg
     1561 atgtcgacga tttgccgttg cgtttccgtc ctagcttctt ccctttcgct gaaccctcag
     1621 ccgaagttga catcggttgc agcaggaagg gcggtgaact caagctgggc aactacggag
     1681 actggctcga aatcctgggc tgcggtatgg tgcatcccaa tgtcctggaa gcctgcggta
     1741 tcgactccac caagtaccag ggcttcggct tcgggatggg aatcgagcgc gtggccatgc
     1801 tgaagtacgg gatccctgac ctgcgcacct tcttcgaagc tgacctccgg tggttgaaac
     1861 attattaagg cgcgccccgt aggcggtcac gactttgcga agcaaagtct agtgagtata
     1921 ctcaagcatt gagtggcccg ccggaggcac cgccttgcgc tgcccccgtc gagccggttg
     1981 gacaccaaaa gggagggtac gtaatgattg aacaagatgg attgcacgca ggttctccgg
     2041 ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg
     2101 atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc
     2161 tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga
     2221 cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc
     2281 tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag
     2341 tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat
     2401 tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg
     2461 tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca
     2521 ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct
     2581 tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg
     2641 gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg
     2701 gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc
     2761 gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat
     2821 gttaattaag attacgccaa gcttgcatgc ctgaggtcga ctctagagga tccttgcccg
     2881 tgttatctca agcggacgac aagaatcgag ggacgggcat gcggcttccc cggctgggcg
     2941 tggcgtttcg catcctggcg ggtctgaccg tgatcgccgg cctgaccgcc gccaccggcg
     3001 tggtcgccgt ctccctcttc ggccgcttcc atgaggggtt cgagcagatc gccacggcca
     3061 aggtgccggg cctcgtcaac gcctcgcagc tggcccagca gagcggcacg ctggccgcca
     3121 acgccccggc cctggtggcg gtgcagagcc agtcggtgcg cgaatcggtg atggcccggc
     3181 tgagcgatca ggtcgccctg ctggaagacc tgatgacccg gctgcgcgac cgcggcgccg
     3241 acatggccga cgtggccgag ttgcagcgcc gcaagaacga gctgatcgcc aacctgatta
     3301 gctaactgaa ctagtcaata cattatcata tcgctatcgt gcgtgatacg acagatggtt
     3361 tggctagctg agtaggtagg accctgaaca cccaggtgga gcgccagctc gccgccgccg
     3421 cggagacggg cgggctggtc ggccgcctga tgcatctggc cgaacgcatc cgcaacgccg
     3481 aggaagcgct gcgggacaag gccaccaccc cccgcaccct ggaggagtgg gaggaggccc
     3541 ggggcgtcga acggcggatc gacctgtgga cggcggaggc gcagcacgcc gtcctgctga
     3601 tgctcgccgc ctcccgcgcc gaccacgagg cgcgggtgga ccggttgcgg caggactacc
     3661 gcgccgcgat ggaccgcgca gcggaggtcg tggccgccct gccctccggc gccgccttcg
     3721 acctggagcc gctgtaccgg gagctggagg cgctggggct cggggaacac aacctgttca
     3781 acagccggct gacggaaatc ggatccccgg gtaccgagct cgaattcatg gcatagatag
     3841 gtgggctgcc cttcctggtt ggcttggttt catcagccat ccgcttgccc tcatctgtta
     3901 cgccggcggt agccggccag cctcgcagag caggattccc gttgagcacc gccaggtgcg
     3961 aataagggac agtgaagaag gaacacccgc tcgcgggtgg gcctacttca cctatcctgc
     4021 ccggctgacg ccgttggata caccaaggaa agtctacacg aaccctttgg caaaatcctg
     4081 tatatcgtgc gaaaaaggat ggatataccg aaaaaatcgc tataatgacc ccgaagcagg
     4141 gttatgcagc ggaaaacgga attcgagcag gtggcacttt tcggggaaat gtgcgcggaa
     4201 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     4261 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg
     4321 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
     4381 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg
     4441 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga
     4501 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc
     4561 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag
     4621 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
     4681 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg
     4741 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
     4801 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt
     4861 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact
     4921 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
     4981 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg
     5041 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
     5101 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac
     5161 tgtcagacca agtttactca tatatacttt a
//
