LOCUS       sequence-386115-        3258 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  terminator from Agrobacterium tumefaciens nopaline synthase (tNOS, 
            MF116010) for T3 position (K/S).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3258)
  AUTHORS   Bonturi N, Pinheiro MJ, de Oliveira PM, Rusadze E, Eichinger T, 
            Liudziute G, De Biaggi JS, Brauer A, Remm M, Miranda EA, 
            Ledesma-Amaro R, Lahtvee PJ
  TITLE     Development of a dedicated Golden Gate Assembly Platform (RtGGA) for
            Rhodotorula toruloides.
  JOURNAL   Metab Eng Commun. 2022 May 23;15:e00200. doi: 
            10.1016/j.mec.2022.e00200. eCollection 2022 Dec.
  PUBMED    35662893
REFERENCE   2  (bases 1 to 3258)
  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"; journalName: "Metab Eng 
            Commun."; date: "2022-05-23"; volume: "15:e00200. doi"; pages: " 
            10.1016/j.mec.2022.e00200. eCollection 2022 Dec"
COMMENT     Sequence Label: (K) tNOS (S)_T3 (SBE120)
FEATURES             Location/Qualifiers
     source          1..3258
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(53..844,845..913))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     676..695
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(914..1018)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1086..1104
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     1410..1429
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1410..1428
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      1489..1740
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     promoter        complement(join(2195..2201,2202..2219,2220..2225))
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    2240..2261
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2381..2398)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2552..3140)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2632..2651)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
       61 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
      121 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
      181 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
      241 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
      301 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
      361 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
      421 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
      481 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
      541 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
      601 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
      661 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
      721 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
      781 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
      841 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
      901 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
      961 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     1021 catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct
     1081 ataaaaatag gcgtatcacg aggccgcccc tgcagccgaa ttatattatt tttgccaaat
     1141 aatttttaac aaaagctctg aagtcttctt catttaaatt cttagatgat acttcatctg
     1201 gaaaattgtc ccaattagta gcatcacgct gtgagtaagt tctaaaccat ttttttattg
     1261 ttgtattatc tctaatctta ctactcgatg agttttcggt attatctcta tttttaactt
     1321 ggagcaggtt ccattcattg tttttttcat catagtgaat aaaatcaact gctttaacac
     1381 ttgtgcctga acaccatatc catccggcgt aatacgactc actataggga gagcggccgc
     1441 cagatcttcc ggatggctcg agtttttcag caagatgatg gtctcagtat cgttcaaaca
     1501 tttggcaata aagtttctta agattgaatc ctgttgccgg tcttgcgatg attatcatat
     1561 aatttctgtt gaattacgtt aagcatgtaa taattaacat gtaatgcatg acgttattta
     1621 tgagatgggt ttttatgatt agagtcccgc aattatacat ttaatacgcg atagaaaaca
     1681 aaatatagcg cgcaaactag gataaattat cgcgcgcggt gtcatctatg ttactagatc
     1741 gggtcgggga gaccggtaga atctttctag aagatctcct acaatattct cagctgccat
     1801 ggaaaatcga tgttcttctt ttattctctc aagattttca ggctgtatat taaaacttat
     1861 attaagaact atgctaacca cctcatcagg aaccgttgta ggtggcgtgg gttttcttgg
     1921 caatcgactc tcatgaaaac tacgagctaa atattcaata tgttcctctt gaccaacttt
     1981 attctgcatt ttttttgaac gaggtttaga gcaagcttca ggaaactgag acaggaattt
     2041 tattaaaaat ttaaattttg aagaaagttc agggttaata gcatccattt tttgctttgc
     2101 aagttcctca gcattcttaa caaaagacgt ctcttttgac atgtttaaag tttaaacctc
     2161 ctgtgtgaaa ttattatccg ctcataattc cacacattat acgagccgga agcataaagt
     2221 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     2281 caattgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     2341 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     2401 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     2461 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     2521 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     2581 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     2641 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     2701 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     2761 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     2821 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     2881 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     2941 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     3001 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     3061 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     3121 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     3181 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     3241 agatcctttt aaattaaa
//
