LOCUS       sequence-208888-        3771 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Provides Nos terminator and 3' part of Tobacco mosaic virus (3NTR, 
            ribozyme) as level 0 GoldenBraid part (GCTT-CGCT).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3771)
  TITLE     Moravec lab plasmids
REFERENCE   2  (bases 1 to 3771)
  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: pUPD2b:TMV-3NTR
FEATURES             Location/Qualifiers
     source          1..3771
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(70..88)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(71..88)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(106..122)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(106..122)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(119..141)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    130..146
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /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)."
     promoter        complement(join(154..160,161..178,179..184))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    199..220
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(337..354)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(508..1096)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(588..607)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1267..2058,2059..2127))
                     /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     1890..1909
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2128..2232)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2310..2765)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2447..2468)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2659..2678
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2891..2913
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2905..2922
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2906..2922
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2929..2948
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2929..2947
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      2978..2987
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      2982..2991
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     terminator      3486..3738
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
ORIGIN
        1 aattcgcggc cgcctgcagg tcgaccatat gggagagctc ccaacgcgtt ggatgcatag
       61 cttgagtatt ctatagtgtc acctaaatag cttggcgtaa tcatggtcat agctgtttcc
      121 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
      181 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
      241 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
      301 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
      361 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
      421 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
      481 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
      541 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
      601 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
      661 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
      721 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
      781 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
      841 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
      901 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
      961 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     1021 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     1081 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     1141 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     1201 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     1261 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     1321 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     1381 tggccccagt gctgcaatga taccgcgcga cccacgctca ccggctccag atttatcagc
     1441 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     1501 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     1561 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     1621 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     1681 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     1741 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     1801 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     1861 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     1921 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     1981 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     2041 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     2101 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     2161 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     2221 aggggttccg cgcacatttc cccgaaaagt gccacctgat gcggtgtgaa ataccgcaca
     2281 gatgcgtaag gagaaaatac cgcatcagga aattgtaagc gttaatattt tgttaaaatt
     2341 cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat
     2401 cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa
     2461 gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg
     2521 cgctggccca ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa
     2581 agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc
     2641 gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag
     2701 tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg
     2761 cgcgtccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct
     2821 tcgctattac gccagctggc gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg
     2881 ccagggtttt cccagtcacg acgttgtaaa acgacggcca gtgaattgta atacgactca
     2941 ctatagggcg aattgggccc gacgtcgcat gctcccggcc gccatggcgg ccgcgggaat
     3001 tcgattgggc gatgagtggt ctcggcttac ctcggtcctg caacttgagg tagtcaagat
     3061 gcataataaa taacggattg tgtccgtaat cacacgtggt gcgtacgata acgcatagtg
     3121 tttttccctc cacttaaatc gaagggttgt gtcttggatc gcgcgggtca aatgtatatg
     3181 gttcatatac atccgcaggc acgtaataaa gcgaggggtt cgaatccccc cgttaccccc
     3241 ggtaggggcc caggtacccg gatgtgtttt ccgggctgat gagtccgtga ggacgaaacc
     3301 tggcctgcag accaatcact agtgcggccc tagaggtgct tggttccatc ctaccacaag
     3361 aacctgttac atttatgttt cttaaaccca gaaagctaaa ctacatgaga agaaacccgc
     3421 ggatcaagga cattttccaa aatgttgcca ttgaaccaag agacatagcc agcgcgaatt
     3481 tccccgatcg ttcaaacatt tggcaataaa gtttcttaag attgaatcct gttgccggtc
     3541 ttgcgatgat tatcatataa tttctgttga attacgttaa gcatgtaata attaacatgt
     3601 aatgcatgac gttatttatg agatgggttt ttatgattag agtcccgcaa ttatacattt
     3661 aatacgcgat agaaaacaaa atatagcgcg caaactagga taaattatcg cgcgcggtgt
     3721 catctatgtt actagatccg ctcgagacca ctcatcgccc aatcactagt g
//
