LOCUS       sequence-386114-        3257 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  terminator from Agrobacterium tumefaciens nopaline synthase (tNOS, 
            MF116010) for T2 position (H/I).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3257)
  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 3257)
  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: (H) tNOS (I) _T2 (SBE119)
FEATURES             Location/Qualifiers
     source          1..3257
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(47..838,839..907))
                     /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     670..689
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(908..1012)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1080..1098
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     1404..1423
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1404..1422
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      complement(1489..1741)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     promoter        complement(join(2188..2194,2195..2212,2213..2218))
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    2233..2254
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2374..2391)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2545..3133)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2625..2644)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
       61 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
      121 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
      181 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
      241 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
      301 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
      361 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
      421 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
      481 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
      541 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
      601 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
      661 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
      721 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
      781 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
      841 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
      901 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
      961 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     1021 cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa
     1081 ataggcgtat cacgaggccg cccctgcagc cgaattatat tatttttgcc aaataatttt
     1141 taacaaaagc tctgaagtct tcttcattta aattcttaga tgatacttca tctggaaaat
     1201 tgtcccaatt agtagcatca cgctgtgagt aagttctaaa ccattttttt attgttgtat
     1261 tatctctaat cttactactc gatgagtttt cggtattatc tctattttta acttggagca
     1321 ggttccattc attgtttttt tcatcatagt gaataaaatc aactgcttta acacttgtgc
     1381 ctgaacacca tatccatccg gcgtaatacg actcactata gggagagcgg ccgccagatc
     1441 ttccggatgg ctcgagtttt tcagcaagat atgcggtctc atgaccccga tctagtaaca
     1501 tagatgacac cgcgcgcgat aatttatcct agtttgcgcg ctatattttg ttttctatcg
     1561 cgtattaaat gtataattgc gggactctaa tcataaaaac ccatctcata aataacgtca
     1621 tgcattacat gttaattatt acatgcttaa cgtaattcaa cagaaattat atgataatca
     1681 tcgcaagacc ggcaacagga ttcaatctta agaaacttta ttgccaaatg tttgaacgat
     1741 cctgagacca tgcatctttc tagaagatct cctacaatat tctcagctgc catggaaaat
     1801 cgatgttctt cttttattct ctcaagattt tcaggctgta tattaaaact tatattaaga
     1861 actatgctaa ccacctcatc aggaaccgtt gtaggtggcg tgggttttct tggcaatcga
     1921 ctctcatgaa aactacgagc taaatattca atatgttcct cttgaccaac tttattctgc
     1981 attttttttg aacgaggttt agagcaagct tcaggaaact gagacaggaa ttttattaaa
     2041 aatttaaatt ttgaagaaag ttcagggtta atagcatcca ttttttgctt tgcaagttcc
     2101 tcagcattct taacaaaaga cgtctctttt gacatgttta aagtttaaac ctcctgtgtg
     2161 aaattattat ccgctcataa ttccacacat tatacgagcc ggaagcataa agtgtaaagc
     2221 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgccaattgc
     2281 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
     2341 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     2401 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     2461 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     2521 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     2581 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     2641 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     2701 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     2761 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     2821 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     2881 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     2941 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     3001 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     3061 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     3121 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     3181 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     3241 tttaaattaa aaatgaa
//
