LOCUS       sequence-491679-        5334 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid for inducible expression of six tRNA encoding genes (argX, 
            glyT, leuW, proL, argU, and ileX) in Escherichia coli, based on 
            pSEVA121 backbone.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5334)
  TITLE     Norholm lab plasmids 2025
REFERENCE   2  (bases 1 to 5334)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pCODE-3
FEATURES             Location/Qualifiers
     source          1..5334
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     9..31
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     misc_feature    56..112
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     terminator      155..249
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     promoter        322..413
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(414..482,483..1274)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     terminator      1278..1317
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     oriT            1464..1572
                     /note="incP origin of transfer"
     CDS             complement(1633..2781)
                     /codon_start=1
                     /product="trans-acting replication protein that binds to 
                     and activates oriV"
                     /label=trfA
                     /translation="MNRTFDRKAYRQELIDAGFSAEDAETIASRTVMRAPRETFQSVGS
                     IVQQATAKIERDSVQLAPPALPAPSAAVERSRRLEQEAAGLAKSMTIDTRGTMTTKKRK
                     TAGEDLAKQVSEAKQAALLKHTKQQIKEMQLSLFDIAPWPDTMRAMPNDTARSALFTTR
                     NKKIPREALQNKVIFHVNKDVKITYTGVELRADDDELVWQQVLEYAKRTPIGEPITFTF
                     YELCQDLGWSINGRYYTKAEECLSRLQATAMGFTSDRVGHLESVSLLHRFRVLDRGKKT
                     SRCQVLIDEEIVVLFAGDHYTKFIWEKYRKLSPTARRMFDYFSSHREPYPLKLETFRLM
                     CGSDSTRVKKWREQVGEACEELRGSGLVEHAWVNDDLVHCKR"
     rep_origin      3174..3705
                     /label=oriV
                     /note="incP origin of replication"
     CDS             complement(3852..4478)
                     /codon_start=1
                     /gene="tetR"
                     /product="tetracycline repressor TetR"
                     /label=TetR
                     /note="TetR binds to the tetracycline operator tetO to 
                     inhibit transcription. This inhibition can be relieved by 
                     adding tetracycline or doxycycline."
                     /translation="MMSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWH
                     VKNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGT
                     RPTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPT
                     TDSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     primer_bind     4383..4402
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     promoter        4494..4549
                     /gene="tetR"
                     /label=tetR/tetA promoters
                     /note="overlapping promoters for bacterial tetR and tetA"
     protein_bind    4530..4548
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     tRNA            5038..5114
                     /gene="argU"
                     /product="E. coli tRNA-Arg recognizing AGA and AGG"
                     /label=argU
     tRNA            5131..5206
                     /gene="ileX"
                     /product="E. coli tRNA-Ile recognizing AUA"
                     /label=ileX
     terminator      complement(5242..5328)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 ttaattaaag cggataacaa tttcacacag gaggccgcct aggccgcggc cgcgcgaatt
       61 cgagctcggt acccggggat cctctagagt cgacctgcag gcatgcaagc ttgcggccgc
      121 gtcgtgactg ggaaaaccct ggcgactagt cttggactcc tgttgataga tccagtaatg
      181 acctcagaac tccatctgga tttgttcaga acgctcggtt gccgccgggc gttttttatt
      241 ggtgagaatc caggggtccc caataattac gatttaaatt agtagcccgc ctaatgagcg
      301 ggcttttttt taattcccct atttgtttat ttttctaaat acattcaaat atgtatccgc
      361 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagca
      421 ttcagcattt tcgtgtggcg ctgattccgt tttttgcggc gttttgcctg ccggtgtttg
      481 cgcatccgga aaccctggtg aaagtgaaag atgcggaaga tcaactgggt gcgcgcgtgg
      541 gctatattga actggatctg aacagcggca aaattctgga atcttttcgt ccggaagaac
      601 gttttccgat gatgagcacc tttaaagtgc tgctgtgcgg tgcggttctg agccgtgtgg
      661 atgcgggcca ggaacaactg ggccgtcgta ttcattatag ccagaacgat ctggtggaat
      721 atagcccggt gaccgaaaaa catctgaccg atggcatgac cgtgcgtgaa ctgtgcagcg
      781 cggcgattac catgagcgat aacaccgcgg cgaacctgct gctgacgacc attggcggtc
      841 cgaaagaact gaccgcgttt ctgcataaca tgggcgatca tgtgacccgt ctggatcgtt
      901 gggaaccgga actgaacgaa gcgattccga acgatgaacg tgataccacc atgccggcag
      961 caatggcgac caccctgcgt aaactgctga cgggtgagct gctgaccctg gcaagccgcc
     1021 agcaactgat tgattggatg gaagcggata aagtggcggg tccgctgctg cgtagcgcgc
     1081 tgccggctgg ctggtttatt gcggataaaa gcggtgcggg cgaacgtggc agccgtggca
     1141 ttattgcggc gctgggcccg gatggtaaac cgagccgtat tgtggtgatt tataccaccg
     1201 gcagccaggc gacgatggat gaacgtaacc gtcagattgc ggaaattggc gcgagcctga
     1261 ttaaacattg gtaaaccgat acaattaaag gctccttttg gagccttttt ttttggacga
     1321 cccttgtcct tttccgctgc ataaccctgc ttcggggtca ttatagcgat tttttcggta
     1381 tatccatcct ttttcgcacg atatacagga ttttgccaaa gggttcgtgt agactttcct
     1441 tggtgtatcc aacggcgtca gccgggcagg ataggtgaag taggcccacc cgcgagcggg
     1501 tgttccttct tcactgtccc ttattcgcac ctggcggtgc tcaacgggaa tcctgctctg
     1561 cgaggctggc cgtaggccgg ccgcgatgca ggtggctgct gaacccccag ccggaactga
     1621 ccccacaagg ccctagcgtt tgcaatgcac caggtcatca ttgacccagg cgtgttccac
     1681 caggccgctg cctcgcaact cttcgcaggc ttcgccgacc tgctcgcgcc acttcttcac
     1741 gcgggtggaa tccgatccgc acatgaggcg gaaggtttcc agcttgagcg ggtacggctc
     1801 ccggtgcgag ctgaaatagt cgaacatccg tcgggccgtc ggcgacagct tgcggtactt
     1861 ctcccatatg aatttcgtgt agtggtcgcc agcaaacagc acgacgattt cctcgtcgat
     1921 caggacctgg caacgggacg ttttcttgcc acggtccagg acgcggaagc ggtgcagcag
     1981 cgacaccgat tccaggtgcc caacgcggtc ggacgtgaag cccatcgccg tcgcctgtag
     2041 gcgcgacagg cattcctcgg ccttcgtgta ataccggcca ttgatcgacc agcccaggtc
     2101 ctggcaaagc tcgtagaacg tgaaggtgat cggctcgccg ataggggtgc gcttcgcgta
     2161 ctccaacacc tgctgccaca ccagttcgtc atcgtcggcc cgcagctcga cgccggtgta
     2221 ggtgatcttc acgtccttgt tgacgtggaa aatgaccttg ttttgcagcg cctcgcgcgg
     2281 gattttcttg ttgcgcgtgg tgaacagggc agagcgggcc gtgtcgtttg gcatcgctcg
     2341 catcgtgtcc ggccacggcg caatatcgaa caaggaaagc tgcatttcct tgatctgctg
     2401 cttcgtgtgt ttcagcaacg cggcctgctt ggcttcgctg acctgttttg ccaggtcctc
     2461 gccggcggtt tttcgcttct tggtcgtcat agttcctcgc gtgtcgatgg tcatcgactt
     2521 cgccaaacct gccgcctcct gttcgagacg acgcgaacgc tccacggcgg ccgatggcgc
     2581 gggcagggca gggggagcca gttgcacgct gtcgcgctcg atcttggccg tagcttgctg
     2641 gactatcgag ccgacggact ggaaggtttc gcggggcgca cgcatgacgg tgcggcttgc
     2701 gatggtttcg gcatcctcgg cggaaaaccc cgcgtcgatc agttcttgcc tgtatgcctt
     2761 ccggtcaaac gtccgattca ttcaccctcc ttgcgggatt gccccggaat taattccccg
     2821 gatcgatccg tcgatcttga tcccctgcgc catcagatcc ttggcggcaa gaaagccatc
     2881 cagtttactt tgcagggctt cccaacctta ccagagggcg ccccagctgg caattccggt
     2941 tcgcttgctg tccataaaac cgcccagtct agctatcgcc atgtaagccc actgcaagct
     3001 acctgctttc tctttgcgct tgcgttttcc cttgtccaga tagcccagta gctgacattc
     3061 atccggggtc agcaccgttt ctgcggactg gctttctacg tggctgccat ttttggggtg
     3121 aggccgttcg cggccgaggg gcgcagcccc tggggggatg ggaggcccgc gttagcgggc
     3181 cgggagggtt cgagaagggg gggcaccccc cttcggcgtg cgcggtcacg cgcacagggc
     3241 gcagccctgg ttaaaaacaa ggtttataaa tattggttta aaagcaggtt aaaagacagg
     3301 ttagcggtgg ccgaaaaacg ggcggaaacc cttgcaaatg ctggattttc tgcctgtgga
     3361 cagcccctca aatgtcaata ggtgcgcccc tcatctgtca gcactctgcc cctcaagtgt
     3421 caaggatcgc gcccctcatc tgtcagtagt cgcgcccctc aagtgtcaat accgcagggc
     3481 acttatcccc aggcttgtcc acatcatctg tgggaaactc gcgtaaaatc aggcgttttc
     3541 gccgatttgc gaggctggcc agctccacgt cgccggccga aatcgagcct gcccctcatc
     3601 tgtcaacgcc gcgccgggtg agtcggcccc tcaagtgtca acgtccgccc ctcatctgtc
     3661 agtgagggcc aagttttccg cgaggtatcc acaacgccgg cggccctaca tggctctgct
     3721 gtagtgagtg ggttgcgctc cggcagcggt cctgatcccc cgcagaaaaa aaggatctca
     3781 agaagatcct ttgatctttt ctacgggaga ccaaaaaaaa aaaaacaccc gttagggtgt
     3841 tttttttttt tttaagaccc actttcacat ttaagttgtt tttctaatcc gcatatgatc
     3901 aattcaaggc cgaataagaa ggctggctct gcaccttggt gatcaaataa ttcgatagct
     3961 tgtcgtaata atggcggcat actatcagta gtaggtgttt ccctttcttc tttagcgact
     4021 tgatgctctt gatcttccaa tacgcaacct aaagtaaaat gccccacagc gctgagtgca
     4081 tataatgcat tctctagtga aaaaccttgt tggcataaaa aggctaattg attttcgaga
     4141 gtttcatact gtttttctgt aggccgtgta cctaaatgta cttttgctcc atcgcgatga
     4201 cttagtaaag cacatctaaa acttttagcg ttattacgta aaaaatcttg ccagctttcc
     4261 ccttctaaag ggcaaaagtg agtatggtgc ctatctaaca tctcaatggc taaggcgtcg
     4321 agcaaagccc gcttattttt tacatgccaa tacaatgtag gctgctctac acctagcttc
     4381 tgggcgagtt tacgggttgt taaaccttcg attccgacct cattaagcag ctctaatgcg
     4441 ctgttaatca ctttactttt atctaatcta gacatcatta attcctaatt tttgttgaca
     4501 ctctatcgtt gatagagtta ttttaccact ccctatcagt gatagagaaa agaattaacg
     4561 gcgctaagcg cccgtagctc agctggatag agcgctgccc tccggaggca gaggtctcag
     4621 gttcgaatcc tgtcgggcgc gccatttagt cccggcgctt gagctgcggt ggtagtaata
     4681 ccgcgtaaca agatttgtag tggcgggcat cgtataatgg ctattacctc agccttccaa
     4741 gctgatgatg cgggttcgat tcccgctgcc cgctccatta ttagaagttg tgacaatgcg
     4801 ggagtggcga aattggtaga cgcaccagat ttaggttctg gcgccgcaag gtgtgcgagt
     4861 tcaagtctcg cctcccgcac cacgacttta aagaattgaa ctaaaaattc aaaaagcagt
     4921 atttcagcca cgagtcggca cgtagcgcag cctggtagcg caccgtcatg gggtgtcggg
     4981 ggtcggaggt tcaaatcctc tcgtgccgac caaaaatcac cgcattgcgg tatcaacgcg
     5041 cccttagctc agttggatag agcaacgacc ttctaagtcg tgggccgcag gttcgaatcc
     5101 tgcagggcgc gccattacaa ttcaatcagt ggccccttag ctcagtggtt agagcaggcg
     5161 actcataatc gcttggtcgc tggttcaagt ccagcagggg ccaccagata tagcggcgcg
     5221 cccagctgtc tagggcggcg gatttgtcct actcaggaga gcgttcaccg acaaacaaca
     5281 gataaaacga aaggcccagt ctttcgactg agcctttcgt tttatttgat gcct
//
