LOCUS       sequence-335111-        3352 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Phytobrick - pNOS.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3352)
  AUTHORS   Gonzalez-Grandio E, Demirer GS, Ma W, Brady S, Landry MP
  TITLE     A Ratiometric Dual Color Luciferase Reporter for Fast 
            Characterization of Transcriptional Regulatory Elements in Plants.
  JOURNAL   ACS Synth Biol. 2021 Sep 14. doi: 10.1021/acssynbio.1c00248.
  PUBMED    34520169
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"; journalName: "ACS Synth 
            Biol. 2021 Sep 14. doi: 10.1021/acssynbio.1c00248."
COMMENT     Sequence Label: pUPD+pNOS
FEATURES             Location/Qualifiers
     source          1..3352
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      30..39
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      34..43
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(73..92)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(74..92)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(99..116)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(99..115)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(108..130)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      256..711
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(343..362)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     553..574
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        789..893
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(894..962,963..1754)
                     /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     complement(1112..1131)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1925..2513
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2414..2433
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2667..2684
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2801..2822
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2837..2842,2843..2860,2861..2867)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2875..2891
                     /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)."
     primer_bind     2880..2902
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2899..2915
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2899..2915
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        2933..2951
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     2933..2950
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     promoter        3132..3311
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
ORIGIN
        1 accactcatc gcccaatcga attcccgcgg ccgccatggc ggccgggagc atgcgacgtc
       61 gggcccaatt cgccctatag tgagtcgtat tacaattcac tggccgtcgt tttacaacgt
      121 cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca tccccctttc
      181 gccagctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc
      241 ctgaatggcg aatggacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
      301 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc
      361 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
      421 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg
      481 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
      541 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct
      601 attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga
      661 tttaacaaaa atttaacgcg aattttaaca aaatattaac gcttacaatt tcctgatgcg
      721 gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atcaggtggc acttttcggg
      781 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
      841 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
      901 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
      961 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
     1021 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     1081 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
     1141 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     1201 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     1261 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
     1321 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     1381 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     1441 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
     1501 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     1561 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     1621 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     1681 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     1741 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
     1801 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     1861 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     1921 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     1981 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     2041 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     2101 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     2161 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     2221 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     2281 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     2341 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     2401 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     2461 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     2521 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     2581 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg
     2641 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc
     2701 caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca
     2761 ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt tagctcactc
     2821 attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt ggaattgtga
     2881 gcggataaca atttcacaca ggaaacagct atgaccatga ttacgccaag ctatttaggt
     2941 gacactatag aatactcaag ctatgcatcc aacgcgttgg gagctctccc atatggtcga
     3001 cctgcaggcg gccgcgaatt cactagtgat tgggcgatga gtggtctcgg gagatacatg
     3061 agaattaagg gagtcacggt atgacccccg ccgatgacgc gggacaagcc gttttacgtt
     3121 tggaactgac agaaccgcaa cgttgaagga gccactgagc cgcgggtttc tggagtttaa
     3181 tgagctaagc acatacgtca gaaaccatta ttgcgcgttc aaaagtcgcc taaggtcact
     3241 atcagctagc aaatatttct tgtcaaaaat gctccactga cgttccataa attcccctcg
     3301 gtatccaatt agagtctcat attcactctc aactcgatcg aggcaatgcg ag
//
