LOCUS       sequence-125715-        4441 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Provides the CDS of the Tobacco echt virus HcPro protein as a level 
            0 GoldenBraid part.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4441)
  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 4441)
  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: pHcPro (GB0067)
FEATURES             Location/Qualifiers
     source          1..4441
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(1491..1509)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1492..1509)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1527..1543)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1527..1543)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1540..1562)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1551..1567
                     /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(1575..1581,1582..1599,1600..1605))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1620..1641
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1758..1775)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1929..2517)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2009..2028)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2688..3479,3480..3548))
                     /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     3311..3330
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3549..3653)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3731..4186)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3868..3889)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4080..4099
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4312..4334
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4326..4343
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4327..4343
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4350..4369
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4350..4368
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      4399..4408
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      4403..4412
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 ctcgaagctc atccaacatt aaaagttttc atttcggatt ccaaacctga atgaacgaat
       61 tcaattagct gggatgttgt gttcattttc aacatgtggt atcctgtcgt tctagaccca
      121 tacgaatcca aaacatgcat tgttttgttg tcatgatcaa ccaaaattct gggtaattca
      181 gcactcagga catctgggta aagaatggaa agtaagtagc atgcagttgc aacatcttgc
      241 attgttggcc acgctccaag ctttggaaca attgtgtccc ttataaactt ggtgaagtcc
      301 tttgcatcct cttccttgac attcactagt agagcaaaga aaatgttcat gtagcaataa
      361 ccttcattag ctatatacat tttctcttca ttgagaactg gaaggtctag gtactttgaa
      421 tcgccagagt tgccaatgac cagatgtctc ttcgttggat gctttagatc cgaatattga
      481 gccttaccat cttcaagagt aacacaacaa catgggtaca cgtaattacc attccgcatt
      541 gaaatgcagt gattcccaat ttctttacgc tcaatagttt cgccttgaat ttgttgcctg
      601 agcgtctgga agtcagttga cattatcaaa ttgccaattg ctagctttct gctacccctg
      661 atatgtttcc tgataacata cttatcgtat ccttcatttg ggtcaatctc agtgaaaaat
      721 cctttaagaa acctctttgc gtgtgcaccc cttagtcccc aaataaaatt cccattctga
      781 tcaagttgat tatcacacat gagtgcgtta ttcacatggg ccttcgatga gattttattt
      841 ctgaaagaac caaggtggcc aatgcgcata ttttcagtgc gattgttcaa gaaccttgct
      901 atttcaagca tatgtgtgct tgcctcttcg agatcggccc ctgttagttt attgccttta
      961 aacagaattt cgctgacagc cagtacgtgt gtgaatggag cttgtttgcg gtcaccaatg
     1021 agttgtttga cgctcacgca ggctgtcaaa tttgtattta ctagtgattt ctgttgcaaa
     1081 aaccttgtga gtagtttctc ggccattggg aaatctggat actgttcttt cagcattgct
     1141 aggagcttac cttggttggt gacactgtct cccgactcac cctcaacaat gtcaggtgtt
     1201 tcaaccatgc aacgtttgca tgtgatctta ccacacggtg aaagtgcttg tgtcaggatt
     1261 gcagccacct caccgcaccg ctcaactgat actcctcttg tacactcatg ggaaattcca
     1321 tctggtctta actccaagaa tttcttagag tatggtatga agaatgcctc agagattgat
     1381 ttgtcgctca ttcgagacca ctcatcgccc aatcactagt gaattcgcgg ccgcctgcag
     1441 gtcgaccata tgggagagct cccaacgcgt tggatgcata gcttgagtat tctatagtgt
     1501 cacctaaata gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg
     1561 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
     1621 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     1681 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     1741 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     1801 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     1861 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     1921 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     1981 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     2041 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     2101 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     2161 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     2221 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     2281 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     2341 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     2401 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     2461 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     2521 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     2581 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     2641 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     2701 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     2761 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     2821 ataccgcgcg acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     2881 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     2941 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     3001 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     3061 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     3121 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     3181 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     3241 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     3301 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     3361 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     3421 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     3481 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     3541 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     3601 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     3661 ccccgaaaag tgccacctga tgcggtgtga aataccgcac agatgcgtaa ggagaaaata
     3721 ccgcatcagg aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa
     3781 atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa
     3841 tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac
     3901 gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgctggccc actacgtgaa
     3961 ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct
     4021 aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa
     4081 gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc
     4141 gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtccat tcgccattca
     4201 ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg
     4261 cgaaaggggg atgtgctgca aggcgattaa gttgggtaac gccagggttt tcccagtcac
     4321 gacgttgtaa aacgacggcc agtgaattgt aatacgactc actatagggc gaattgggcc
     4381 cgacgtcgca tgctcccggc cgccatggcg gccgcgggaa ttcgattggg cgatgagtgg
     4441 t
//
