LOCUS       sequence-224424-        4997 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of CURT_fluoB controlled by the ARA promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4997)
  AUTHORS   Behrendorff JBYH, Sandoval-Ibanez OA, Sharma A, Pribil M
  TITLE     Membrane-bound protein scaffolding in diverse hosts using thylakoid 
            protein CURT1A.
  JOURNAL   ACS Synth Biol. 2019 Mar 18. doi: 10.1021/acssynbio.8b00418.
  PUBMED    30884945
REFERENCE   2  (bases 1 to 4997)
  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. 2019 Mar 18. doi: 10.1021/acssynbio.8b00418."
COMMENT     Sequence Label: pBAD/CURT_fluoB
FEATURES             Location/Qualifiers
     source          1..4997
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(19..38)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     179..201
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(357..1235)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        1262..1546
                     /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     1469..1488
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             join(1904..1906,1907..2620)
                     /codon_start=1
                     /product="enhanced YFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEYFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGLMCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(1946..1967)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2207..2226)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2554..2575
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2659..2676)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      2829..2915
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3007..3034
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        3053..3144
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3145..3213,3214..4005)
                     /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(3363..3382)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4176..4764
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4665..4684
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4918..4935
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    join(4950..4997,1..93)
                     /label=bom
                     /note="basis of mobility region from pBR322"
ORIGIN
        1 catatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc agtatacact
       61 ccgctatcgc tacgtgactg ggtcatggct gcgccccgac acccgccaac acccgctgac
      121 gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc
      181 gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag gcagcagatc
      241 aattcgcgcg cgaaggcgaa gcggcatgca taatgtgcct gtcaaatgga cgaagcaggg
      301 attctgcaaa ccctatgcta ctccgtcaag ccgtcaattg tctgattcgt taccaattat
      361 gacaacttga cggctacatc attcactttt tcttcacaac cggcacggaa ctcgctcggg
      421 ctggccccgg tgcatttttt aaatacccgc gagaaataga gttgatcgtc aaaaccaaca
      481 ttgcgaccga cggtggcgat aggcatccgg gtggtgctca aaagcagctt cgcctggctg
      541 atacgttggt cctcgcgcca gcttaagacg ctaatcccta actgctggcg gaaaagatgt
      601 gacagacgcg acggcgacaa gcaaacatgc tgtgcgacgc tggcgatatc aaaattgctg
      661 tctgccaggt gatcgctgat gtactgacaa gcctcgcgta cccgattatc catcggtgga
      721 tggagcgact cgttaatcgc ttccatgcgc cgcagtaaca attgctcaag cagatttatc
      781 gccagcagct ccgaatagcg cccttcccct tgcccggcgt taatgatttg cccaaacagg
      841 tcgctgaaat gcggctggtg cgcttcatcc gggcgaaaga accccgtatt ggcaaatatt
      901 gacggccagt taagccattc atgccagtag gcgcgcggac gaaagtaaac ccactggtga
      961 taccattcgc gagcctccgg atgacgaccg tagtgatgaa tctctcctgg cgggaacagc
     1021 aaaatatcac ccggtcggca aacaaattct cgtccctgat ttttcaccac cccctgaccg
     1081 cgaatggtga gattgagaat ataacctttc attcccagcg gtcggtcgat aaaaaaatcg
     1141 agataaccgt tggcctcaat cggcgttaaa cccgccacca gatgggcatt aaacgagtat
     1201 cccggcagca ggggatcatt ttgcgcttca gccatacttt tcatactccc gccattcaga
     1261 gaagaaacca attgtccata ttgcatcaga cattgccgtc actgcgtctt ttactggctc
     1321 ttctcgctaa ccaaaccggt aaccccgctt attaaaagca ttctgtaaca aagcgggacc
     1381 aaagccatga caaaaacgcg taacaaaagt gtctataatc acggcagaaa agtccacatt
     1441 gattatttgc acggcgtcac actttgctat gccatagcat ttttatccat aagattagcg
     1501 gatcctacct gacgcttttt atcgcaactc tctactgttt ctccataccc gttttttggg
     1561 ctaacaggag gaattaacca tggcttcttc agaagagacc tcgtccattg ataccaacga
     1621 actcatcaca gatttgaagg aaaagtggga tggtcttgag aacaaatcta ctgtacttat
     1681 atatggagga ggagccattg ttgctgtttg gttatcttcc attgttgttg gtgccatcaa
     1741 ctctgttcct ctgcttccga aagttatgga acttgtcgga ctagggtaca ctggatggtt
     1801 tgtctacaga taccttctct tcaagtcaag cagaaaggaa ttggctgagg atattgaatc
     1861 cttgaagaag aagatcgcag gaagcgaagg aggtggtgga tctgtgagca agggcgagga
     1921 gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa
     1981 gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagtt
     2041 catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccttcggcta
     2101 cggcctgatg tgcttcgccc gctaccccga ccacatgaag cagcacgact tcttcaagtc
     2161 cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta
     2221 caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa
     2281 gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa
     2341 cagccacaac gtctatatca tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa
     2401 gatccgccac aacatcgagg acggcagcgt gcagctcgcc gaccactacc agcagaacac
     2461 ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagct accagtccaa
     2521 gctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc
     2581 cgccgggatc actctcggca tggacgagct gtacaagtaa aagcttggct gttttggcgg
     2641 atgagagaag attttcagcc tgatacagat taaatcagaa cgcagaagcg gtctgataaa
     2701 acagaatttg cctggcggca gtagcgcggt ggtcccacct gaccccatgc cgaactcaga
     2761 agtgaaacgc cgtagcgccg atggtagtgt ggggtctccc catgcgagag tagggaactg
     2821 ccaggcatca aataaaacga aaggctcagt cgaaagactg ggcctttcgt tttatctgtt
     2881 gtttgtcggt gaacgctctc ctgagtagga caaatccgcc gggagcggat ttgaacgttg
     2941 cgaagcaacg gcccggaggg tggcgggcag gacgcccgcc ataaactgcc aggcatcaaa
     3001 ttaagcagaa ggccatcctg acggatggcc tttttgcgtt tctacaaact cttttgttta
     3061 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     3121 caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc
     3181 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa
     3241 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt
     3301 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt
     3361 ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc
     3421 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg
     3481 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg
     3541 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac
     3601 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca
     3661 aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta
     3721 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat
     3781 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa
     3841 tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag
     3901 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat
     3961 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt
     4021 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     4081 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     4141 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     4201 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     4261 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     4321 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     4381 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     4441 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     4501 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     4561 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     4621 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     4681 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     4741 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     4801 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     4861 cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc
     4921 gagtcagtga gcgaggaagc ggaagagcgc ctgatgcggt attttctcct tacgcatctg
     4981 tgcggtattt cacaccg
//
