LOCUS       sequence-229431-        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"
     CDS             join(358..360,361..1074)
                     /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(400..421)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(661..680)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1008..1029
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1113..1130)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1283..1369
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1461..1488
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        1507..1598
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1599..1667,1668..2459)
                     /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(1817..1836)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2630..3218
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3119..3138
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3372..3389
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3404..3544
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3470..3489)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3630..3652
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3808..4686)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        4713..4997
                     /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     4920..4939
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
ORIGIN
        1 acccgttttt tgggctaaca ggaggaatta accatggctt cttcagaaga aacatcgtcc
       61 attgatacca acgaactcat cacagatttg aaggaaaagt gggatggtct tgagaacaaa
      121 tctactgtac ttatatatgg aggaggagcc attgttgctg tttggttatc ttccattgtt
      181 gttggtgcca tcaactctgt tcctctgctt ccgaaagtta tggaacttgt cggactaggg
      241 tacactggat ggtttgtcta cagatacctt ctcttcaagt caagcagaaa ggaattggct
      301 gaggatattg aatccttgaa gaagaagatc gcaggaagcg aaggaggtgg tggatctgtg
      361 agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac
      421 gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag
      481 ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg
      541 accaccttcg gctacggcct gatgtgcttc gcccgctacc ccgaccacat gaagcagcac
      601 gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag
      661 gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac
      721 cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg
      781 gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa gaacggcatc
      841 aaggtgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct cgccgaccac
      901 taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg
      961 agctaccagt ccaagctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg
     1021 gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa gtaaaagctt
     1081 ggctgttttg gcggatgaga gaagattttc agcctgatac agattaaatc agaacgcaga
     1141 agcggtctga taaaacagaa tttgcctggc ggcagtagcg cggtggtccc acctgacccc
     1201 atgccgaact cagaagtgaa acgccgtagc gccgatggta gtgtggggtc tccccatgcg
     1261 agagtaggga actgccaggc atcaaataaa acgaaaggct cagtcgaaag actgggcctt
     1321 tcgttttatc tgttgtttgt cggtgaacgc tctcctgagt aggacaaatc cgccgggagc
     1381 ggatttgaac gttgcgaagc aacggcccgg agggtggcgg gcaggacgcc cgccataaac
     1441 tgccaggcat caaattaagc agaaggccat cctgacggat ggcctttttg cgtttctaca
     1501 aactcttttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     1561 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg
     1621 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
     1681 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg
     1741 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga
     1801 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc
     1861 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag
     1921 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
     1981 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg
     2041 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
     2101 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt
     2161 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact
     2221 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
     2281 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg
     2341 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
     2401 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac
     2461 tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat ttttaattta
     2521 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
     2581 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     2641 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     2701 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     2761 agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     2821 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     2881 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     2941 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     3001 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     3061 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     3121 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     3181 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     3241 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg
     3301 attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa
     3361 cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg cggtattttc
     3421 tccttacgca tctgtgcggt atttcacacc gcatatggtg cactctcagt acaatctgct
     3481 ctgatgccgc atagttaagc cagtatacac tccgctatcg ctacgtgact gggtcatggc
     3541 tgcgccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc
     3601 atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc
     3661 gtcatcaccg aaacgcgcga ggcagcagat caattcgcgc gcgaaggcga agcggcatgc
     3721 ataatgtgcc tgtcaaatgg acgaagcagg gattctgcaa accctatgct actccgtcaa
     3781 gccgtcaatt gtctgattcg ttaccaatta tgacaacttg acggctacat cattcacttt
     3841 ttcttcacaa ccggcacgga actcgctcgg gctggccccg gtgcattttt taaatacccg
     3901 cgagaaatag agttgatcgt caaaaccaac attgcgaccg acggtggcga taggcatccg
     3961 ggtggtgctc aaaagcagct tcgcctggct gatacgttgg tcctcgcgcc agcttaagac
     4021 gctaatccct aactgctggc ggaaaagatg tgacagacgc gacggcgaca agcaaacatg
     4081 ctgtgcgacg ctggcgatat caaaattgct gtctgccagg tgatcgctga tgtactgaca
     4141 agcctcgcgt acccgattat ccatcggtgg atggagcgac tcgttaatcg cttccatgcg
     4201 ccgcagtaac aattgctcaa gcagatttat cgccagcagc tccgaatagc gcccttcccc
     4261 ttgcccggcg ttaatgattt gcccaaacag gtcgctgaaa tgcggctggt gcgcttcatc
     4321 cgggcgaaag aaccccgtat tggcaaatat tgacggccag ttaagccatt catgccagta
     4381 ggcgcgcgga cgaaagtaaa cccactggtg ataccattcg cgagcctccg gatgacgacc
     4441 gtagtgatga atctctcctg gcgggaacag caaaatatca cccggtcggc aaacaaattc
     4501 tcgtccctga tttttcacca ccccctgacc gcgaatggtg agattgagaa tataaccttt
     4561 cattcccagc ggtcggtcga taaaaaaatc gagataaccg ttggcctcaa tcggcgttaa
     4621 acccgccacc agatgggcat taaacgagta tcccggcagc aggggatcat tttgcgcttc
     4681 agccatactt ttcatactcc cgccattcag agaagaaacc aattgtccat attgcatcag
     4741 acattgccgt cactgcgtct tttactggct cttctcgcta accaaaccgg taaccccgct
     4801 tattaaaagc attctgtaac aaagcgggac caaagccatg acaaaaacgc gtaacaaaag
     4861 tgtctataat cacggcagaa aagtccacat tgattatttg cacggcgtca cactttgcta
     4921 tgccatagca tttttatcca taagattagc ggatcctacc tgacgctttt tatcgcaact
     4981 ctctactgtt tctccat
//
