LOCUS       sequence-319283-        3352 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Nos promoter for N-terminal fusions.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3352)
  TITLE     Orzaez lab plasmids
REFERENCE   2  (bases 1 to 3352)
  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: pUPD Pnos (GB0555)
FEATURES             Location/Qualifiers
     source          1..3352
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(46..225)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     promoter        complement(406..424)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(407..424)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(442..458)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(442..458)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(455..477)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    466..482
                     /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(490..496,497..514,515..520))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    535..556
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(673..690)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(844..1432)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(924..943)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1603..2394,2395..2463))
                     /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     2226..2245
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2464..2568)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2646..3101)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2783..2804)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2995..3014
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3227..3249
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3241..3258
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3242..3258
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3265..3284
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3265..3283
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      3314..3323
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      3318..3327
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 tggtctcgat gggcctcgat cgagttgaga gtgaatatga gactctaatt ggataccgag
       61 gggaatttat ggaacgtcag tggagcattt ttgacaagaa atatttgcta gctgatagtg
      121 accttaggcg acttttgaac gcgcaataat ggtttctgac gtatgtgctt agctcattaa
      181 actccagaaa cccgcggctc agtggctcct tcaacgttgc ggttctgtca gttccaaacg
      241 taaaacggct tgtcccgcgt catcggcggg ggtcataccg tgactccctt aattctcatg
      301 tatctcccga gaccactcat cgcccaatca ctagtgaatt cgcggccgcc tgcaggtcga
      361 ccatatggga gagctcccaa cgcgttggat gcatagcttg agtattctat agtgtcacct
      421 aaatagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
      481 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
      541 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
      601 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
      661 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
      721 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
      781 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
      841 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
      901 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
      961 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     1021 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     1081 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     1141 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     1201 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     1261 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     1321 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     1381 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     1441 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     1501 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     1561 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     1621 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     1681 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     1741 gcgcgaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     1801 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     1861 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     1921 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     1981 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     2041 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     2101 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     2161 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     2221 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     2281 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     2341 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     2401 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     2461 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     2521 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     2581 aaaagtgcca cctgatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca
     2641 tcaggaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag
     2701 ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac
     2761 cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga
     2821 ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgct ggcccactac gtgaaccatc
     2881 accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg
     2941 gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa
     3001 gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac
     3061 caccacaccc gccgcgctta atgcgccgct acagggcgcg tccattcgcc attcaggctg
     3121 cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa
     3181 gggggatgtg ctgcaaggcg attaagttgg gtaacgccag ggttttccca gtcacgacgt
     3241 tgtaaaacga cggccagtga attgtaatac gactcactat agggcgaatt gggcccgacg
     3301 tcgcatgctc ccggccgcca tggcggccgc gggaattcga ttgggcgatg ag
//
