LOCUS       sequence-394143-        5634 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Golden Gate assembly of expression constructs containing pBAD 
            promoter (carbenicillin resistance).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5634)
  AUTHORS   Lesley Milnes,  Mark Youles,  Josephine H. R. Maidment
  TITLE     pOPARA: Vectors for Golden Gate assembly of expression constructs 
            containing the araBAD promoter
  JOURNAL   bioRxiv
REFERENCE   2  (bases 1 to 5634)
  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: "bioRxiv"
COMMENT     Sequence Label: pOPARA1
FEATURES             Location/Qualifiers
     source          1..5634
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(109..987)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        1014..1298
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     1221..1240
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     protein_bind    1448..1469
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1484..1489,1490..1507,1508..1514)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1522..1538
                     /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)."
     RBS             1556..1567
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             1574..2251
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     terminator      2296..2367
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2383..2410
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     primer_bind     complement(2490..2507)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      2660..2746
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2838..2865
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        2911..2982
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2983..3051,3052..3843)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(3201..3220)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3893..4347
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3979..3998)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     rep_origin      4458..5046
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4947..4966
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5200..5217
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     5383..5405
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5544..5563
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 ttgacagctt atcatcgatg cataatgtgc ctgtcaaatg gacgaagcag ggattctgca
       61 aaccctatgc tactccgtca agccgtcaat tgtctgattc gttaccaatt atgacaactt
      121 gacggctaca tcattcactt tttcttcaca accggcacgg aactcgctcg ggctggcccc
      181 ggtgcatttt ttaaataccc gcgagaaata gagttgatcg tcaaaaccaa cattgcgacc
      241 gacggtggcg ataggcatcc gggtggtgct caaaagcagc ttcgcctggc tgatacgttg
      301 gtcctcgcgc cagcttaaga cgctaatccc taactgctgg cggaaaagat gtgacagacg
      361 cgacggcgac aagcaaacat gctgtgcgac gctggcgata tcaaaattgc tgtctgccag
      421 gtgatcgctg atgtactgac aagcctcgcg tacccgatta tccatcggtg gatggagcga
      481 ctcgttaatc gcttccatgc gccgcagtaa caattgctca agcagattta tcgccagcag
      541 ctccgaatag cgcccttccc cttgcccggc gttaatgatt tgcccaaaca ggtcgctgaa
      601 atgcggctgg tgcgcttcat ccgggcgaaa gaaccccgta ttggcaaata ttgacggcca
      661 gttaagccat tcatgccagt aggcgcgcgg acgaaagtaa acccactggt gataccattc
      721 gcgagcctcc ggatgacgac cgtagtgatg aatctctcct ggcgggaaca gcaaaatatc
      781 acccggtcgg caaacaaatt ctcgtccctg atttttcacc accccctgac cgcgaatggt
      841 gagattgaga atataacctt tcattcccag cggtcggtcg ataaaaaaat cgagataacc
      901 gttggcctca atcggcgtta aacccgccac cagatgggca ttaaacgagt atcccggcag
      961 caggggatca ttttgcgctt cagccatact tttcatactc ccgccattca gagaagaaac
     1021 caattgtcca tattgcatca gacattgccg tcactgcgtc ttttactggc tcttctcgct
     1081 aaccaaaccg gtaaccccgc ttattaaaag cattctgtaa caaagcggga ccaaagccat
     1141 gacaaaaacg cgtaacaaaa gtgtctataa tcacggcaga aaagtccaca ttgattattt
     1201 gcacggcgtc acactttgct atgccatagc atttttatcc ataagattag cggatcctac
     1261 ctgacgcttt ttatcgcaac tctctactgt ttctccatac ccgttttttt gggctagcag
     1321 gaggaattca caatgtgaga ccgatatcaa tacgcaaacc gcctctcccc gcgcgttggc
     1381 cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca
     1441 acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc
     1501 cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacatac tagagaaaga
     1561 ggagaaatac tagatggctt cctccgagga cgttatcaaa gagttcatgc gtttcaaagt
     1621 tcgtatggaa ggttccgtta acggtcacga gttcgaaatc gaaggtgaag gtgaaggtcg
     1681 tccgtacgaa ggtacccaga ccgctaaact gaaagttacc aaaggtggtc cgctgccgtt
     1741 cgcttgggac atcctgtccc cgcagttcca gtacggttcc aaagcttacg ttaaacaccc
     1801 ggctgacatc ccggactacc tgaaactgtc cttcccggaa ggtttcaaat gggaacgtgt
     1861 tatgaacttc gaggacggtg gtgttgttac cgttacccag gactcctccc tgcaagacgg
     1921 tgagttcatc tacaaagtta aactgcgtgg taccaacttc ccgtccgacg gtccggttat
     1981 gcagaaaaaa accatgggtt gggaagcttc caccgaacgt atgtacccgg aggacggtgc
     2041 tctgaaaggt gaaatcaaaa tgcgtctgaa actgaaagac ggtggtcact acgacgctga
     2101 agttaaaacc acctacatgg ctaaaaaacc ggttcagctg ccgggtgctt acaaaaccga
     2161 catcaaactg gacatcacct cccacaacga ggactacacc atcgttgaac agtacgaacg
     2221 tgctgaaggt cgtcactcca ccggtgctta ataacgctga tagtgctagt gtagatcgct
     2281 actagagcca ggcatcaaat aaaacgaaag gctcagtcga aagactgggc ctttcgtttt
     2341 atctgttgtt tgtcggtgaa cgctctctac tagagtcaca ctggctcacc ttcgggtggg
     2401 cctttctgcg tttataggtc tcagctttct agagtcgacc tgcaggcatg caagcttggc
     2461 tgttttggcg gatgagagaa gattttcagc ctgatacaga ttaaatcaga acgcagaagc
     2521 ggtctgataa aacagaattt gcctggcggc agtagcgcgg tggtcccacc tgaccccatg
     2581 ccgaactcag aagtgaaacg ccgtagcgcc gatggtagtg tggggtttcc ccatgcgaga
     2641 gtagggaact gccaggcatc aaataaaacg aaaggctcag tcgaaagact gggcctttcg
     2701 ttttatctgt tgtttgtcgg tgaacgctct cctgagtagg acaaatccgc cgggagcgga
     2761 tttgaacgtt gcgaagcaac ggcccggagg gtggcgggca ggacgcccgc cataaactgc
     2821 caggcatcaa attaagcaga aggccatcct gacggatggc ctttttgcgt ttctacaaac
     2881 tcttttgttt atttttctaa atacaagaga cgttcaaata tgtatccgct catgagacaa
     2941 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
     3001 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
     3061 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
     3121 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
     3181 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtgttga cgccgggcaa
     3241 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
     3301 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
     3361 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
     3421 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
     3481 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca
     3541 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
     3601 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
     3661 tggtttattg ctgataaatc tggagccggt gagcgtggat ctcgcggtat cattgcagca
     3721 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
     3781 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
     3841 taactgtcag accaagttta ctcatatata ctttagattg atttacgtct cgcgcgccct
     3901 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
     3961 ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
     4021 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
     4081 ggcacctcga ccccaaaaaa cttgatttgg gtgatggttc acgtagtggg ccatcgccct
     4141 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
     4201 tccaaacttg aacaacactc aaccctatct cgggctattc ttttgattta taagggattt
     4261 tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
     4321 ttaacaaaat attaacgttt acaatttaaa aggatctagg tgaagatcct ttttgataat
     4381 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     4441 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     4501 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     4561 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg
     4621 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     4681 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     4741 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     4801 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     4861 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     4921 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     4981 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     5041 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     5101 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag
     5161 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa
     5221 gcggaagagc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc
     5281 atagggtcat ggctgcgccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt
     5341 gtctgctccc ggcatccgct tacagacaag ctgtgaccga ctccgggagc tgcatgtgtc
     5401 agaggttttc accgtcatca ccgaaacgcg cgaggcagca aggagatggc gcccaacagt
     5461 cccccggcca cggggcctgc caccataccc acgccgaaac aagcgctcat gagcccgaag
     5521 tggcgagccc gatcttcccc atcggtgatg tcggcgatat aggcgccagc aaccgcacct
     5581 gtggcgccgg tgatgccggc cacgatgcgt ccggcgtaga ggatctgctc atgt
//
