LOCUS       sequence-135572-        3682 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Provides the 5'-untranslated region of the Arabidopsis thaliana 
            actin2 gene as a level 0 GoldenBraid part.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3682)
  AUTHORS   Sarrion-Perdigones A, Vazquez-Vilar M, Palaci J, Castelijns B, 
            Forment J, Ziarsolo P, Blanca J, Granell A, Orzaez D
  TITLE     GoldenBraid 2.0: a comprehensive DNA assembly framework for plant 
            synthetic biology.
  JOURNAL   Plant Physiol. 2013 Jul;162(3):1618-31.
  PUBMED    23669743
REFERENCE   2  (bases 1 to 3682)
  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: "Plant 
            Physiol."; date: "2013-07"; volume: "162(3)"; pages: "1618-31"
COMMENT     Sequence Label: p5'UTRAct2 (GB0193)
FEATURES             Location/Qualifiers
     source          1..3682
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(732..750)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(733..750)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(768..784)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(768..784)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(781..803)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    792..808
                     /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(816..822,823..840,841..846))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    861..882
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(999..1016)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1170..1758)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1250..1269)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1929..2720,2721..2789))
                     /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     2552..2571
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2790..2894)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2972..3427)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3109..3130)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3321..3340
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3553..3575
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3567..3584
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3568..3584
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3591..3610
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3591..3609
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      3640..3649
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      3644..3653
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 ctcgtactat tgtctcgttg tcctcctcac tttcatcagc cgttttgaat ctccggcgac
       61 ttgacagaga agaacaagga agaagactaa gagagaaagt aagagataat ccaggagatt
      121 cattctccgt tttgaatctt cctcaatctc atcttcttcc gctctttctt tccaaggtaa
      181 taggaacttt ctggatctac tttatttgct ggatctcgat cttgttttct caatttcctt
      241 gagatctgga attcgtttaa tttggatctg tgaacctcca ctaaatcttt tggttttact
      301 agaatcgatc taagttgacc gatcagttag ctcgattata gctaccagaa tttggcttga
      361 ccttgatgga gagatccatg ttcatgttac ctgggaaatg atttgtatat gtgaattgaa
      421 atctgaactg ttgaagttag attgaatctg aacactgtca atgttagatt gaatctgaac
      481 actgtttaag gttagatgaa gtttgtgtat agattcttcg aaactttagg atttgtagtg
      541 tcgtacgttg aacagaaagc tatttctgat tcaatcaggg tttatttgac tgtattgaac
      601 tctttttgtg tgtttgcagc tcataaaaaa atgcgagacc actcatcgcc caatcactag
      661 tgaattcgcg gccgcctgca ggtcgaccat atgggagagc tcccaacgcg ttggatgcat
      721 agcttgagta ttctatagtg tcacctaaat agcttggcgt aatcatggtc atagctgttt
      781 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
      841 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
      901 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
      961 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     1021 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     1081 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     1141 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     1201 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     1261 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     1321 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     1381 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     1441 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     1501 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     1561 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     1621 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     1681 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     1741 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     1801 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     1861 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     1921 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     1981 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     2041 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     2101 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     2161 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     2221 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     2281 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     2341 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     2401 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     2461 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     2521 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     2581 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     2641 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     2701 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     2761 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     2821 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     2881 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg atgcggtgtg aaataccgca
     2941 cagatgcgta aggagaaaat accgcatcag gaaattgtaa gcgttaatat tttgttaaaa
     3001 ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa
     3061 atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac
     3121 aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag
     3181 ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt
     3241 aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg
     3301 gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca
     3361 agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag
     3421 ggcgcgtcca ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct
     3481 cttcgctatt acgccagctg gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa
     3541 cgccagggtt ttcccagtca cgacgttgta aaacgacggc cagtgaattg taatacgact
     3601 cactataggg cgaattgggc ccgacgtcgc atgctcccgg ccgccatggc ggccgcggga
     3661 attcgattgg gcgatgagtg gt
//
