LOCUS       sequence-359622-        3433 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  4 copies of the HRE DNA binding motif.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3433)
  AUTHORS   Sarrion-Perdigones A, Gonzalez Y, Venken KJT
  TITLE     Synthetic Assembly DNA Cloning of Multiplex Hextuple Luciferase 
            Reporter Plasmids.
  JOURNAL   Methods Mol Biol. 2022;2524:409-432. doi: 
            10.1007/978-1-0716-2453-1_32.
  PUBMED    35821490
REFERENCE   2  (bases 1 to 3433)
  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: "Methods Mol
            Biol."; date: "2022"; volume: "2524:409-432. doi"; pages: " 
            10.1007/978-1-0716-2453-1_32"
COMMENT     Sequence Label: TB:4xHRE_RE:MiniP
FEATURES             Location/Qualifiers
     source          1..3433
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     138..157
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     370..392
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     384..401
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     385..401
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     408..427
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        408..426
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      457..466
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      461..470
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(540..571)
                     /label=minP
                     /note="minimal TATA-box promoter with low basal activity"
     misc_feature    726..817
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     complement(758..777)
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     polyA_signal    831..879
                     /note="synthetic polyadenylation signal"
     promoter        complement(982..1000)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(983..1000)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1018..1034)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1018..1034)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1031..1053)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1042..1058
                     /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(1066..1072,1073..1090,1091..1096))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1111..1132
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1249..1266)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1420..2008)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1500..1519)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2179..2970,2971..3039))
                     /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     2802..2821
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3040..3144)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(join(3222..3433,1..244))
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3359..3380)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 gctggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa
       61 gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg
      121 aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt
      181 gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc
      241 gcgtccattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt
      301 cgctattacg ccagctggcg aaagggggat gtgctgcaag gcgattaagt tgggtaacgc
      361 cagggttttc ccagtcacga cgttgtaaaa cgacggccag tgaattgtaa tacgactcac
      421 tatagggcga attgggcccg acgtcgcatg ctcccggccg ccatggcggc cgcgggaatt
      481 cgattgggcg atgagtggtc tcgcattggt ggctttacca acagtaccgg attgccaagc
      541 tggaagtcga gcttccatta tataccctct agtgtctaag cttggccgcc gaggccagat
      601 ttgatatcct cgagggacta ggcagcacgt agtcactcta gactaggcag cacgtagtca
      661 cgcggccgcc taggcagcac gtagtcacag atctctaggc agcacgtagt cacctcctga
      721 cagcgagaga aatgttctgg cacctgcact tgcactgggg acagcctatt ttgctagttt
      781 gttttgtttc gttttgtttt gatggagagc gtatgttagt actatcgatt cacacaaaaa
      841 accaacacac agatgtaatg aaaataaaga tattttattc tcccgagacc actcatcgcc
      901 caatcactag tgaattcgcg gccgcctgca ggtcgaccat atgggagagc tcccaacgcg
      961 ttggatgcat agcttgagta ttctatagtg tcacctaaat agcttggcgt aatcatggtc
     1021 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     1081 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     1141 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     1201 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     1261 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     1321 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     1381 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     1441 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     1501 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     1561 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     1621 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     1681 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     1741 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     1801 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     1861 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     1921 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     1981 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     2041 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     2101 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     2161 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     2221 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     2281 gggcttacca tctggcccca gtgctgcaat gataccgcgc gacccacgct caccggctcc
     2341 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     2401 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     2461 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     2521 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     2581 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     2641 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     2701 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     2761 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     2821 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     2881 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     2941 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     3001 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     3061 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     3121 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg atgcggtgtg
     3181 aaataccgca cagatgcgta aggagaaaat accgcatcag gaaattgtaa gcgttaatat
     3241 tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga
     3301 aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc
     3361 agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac
     3421 cgtctatcag ggc
//
