LOCUS       sequence-229425-        4997 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of CURT_fluoA 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_fluoA
FEATURES             Location/Qualifiers
     source          1..4997
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..285
                     /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     208..227
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             join(319..321,322..324,325..1035)
                     /codon_start=1
                     /product="enhanced YFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLMCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(364..385)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(625..644)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     972..993
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1398..1415)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1568..1654
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1746..1773
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        1792..1883
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1884..1952,1953..2744)
                     /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(2102..2121)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2915..3503
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3404..3423
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3657..3674
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3689..3829
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3755..3774)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3915..3937
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4093..4971)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
ORIGIN
        1 aagaaaccaa ttgtccatat tgcatcagac attgccgtca ctgcgtcttt tactggctct
       61 tctcgctaac caaaccggta accccgctta ttaaaagcat tctgtaacaa agcgggacca
      121 aagccatgac aaaaacgcgt aacaaaagtg tctataatca cggcagaaaa gtccacattg
      181 attatttgca cggcgtcaca ctttgctatg ccatagcatt tttatccata agattagcgg
      241 atcctacctg acgcttttta tcgcaactct ctactgtttc tccatacccg ttttttgggc
      301 taacaggagg aattaaccat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc
      361 atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc
      421 gagggcgatg ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg
      481 cccgtgccct ggcccaccct cgtgaccacc ttcggctacg gcctgatgtg cttcgcccgc
      541 taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc
      601 caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag
      661 ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac
      721 ggcaacatcc tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcatg
      781 gccgacaagc agaagaacgg catcaaggtg aacttcaaga tccgccacaa catcgaggac
      841 ggcagcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg
      901 ctgctgcccg acaaccacta cctgagctac cagtccaagc tgagcaaaga ccccaacgag
      961 aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg
     1021 gacgagctgt acaagggagg tggtggatct gcttcttcag aagagacctc gtccattgat
     1081 accaacgaac tcatcacaga tttgaaggaa aagtgggatg gtcttgagaa caaatctact
     1141 gtacttatat atggaggagg agccattgtt gctgtttggt tatcttccat tgttgttggt
     1201 gccatcaact ctgttcctct gcttccgaaa gttatggaac ttgtcggact agggtacact
     1261 ggatggtttg tctacagata ccttctcttc aagtcaagca gaaaggaatt ggctgaggat
     1321 attgaatcct tgaagaagaa gatcgcagga agcgaataga agcttggctg ttttggcgga
     1381 tgagagaaga ttttcagcct gatacagatt aaatcagaac gcagaagcgg tctgataaaa
     1441 cagaatttgc ctggcggcag tagcgcggtg gtcccacctg accccatgcc gaactcagaa
     1501 gtgaaacgcc gtagcgccga tggtagtgtg gggtctcccc atgcgagagt agggaactgc
     1561 caggcatcaa ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt ttatctgttg
     1621 tttgtcggtg aacgctctcc tgagtaggac aaatccgccg ggagcggatt tgaacgttgc
     1681 gaagcaacgg cccggagggt ggcgggcagg acgcccgcca taaactgcca ggcatcaaat
     1741 taagcagaag gccatcctga cggatggcct ttttgcgttt ctacaaactc ttttgtttat
     1801 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     1861 aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct
     1921 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
     1981 atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta
     2041 agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc
     2101 tgctatgtgg cgcggtatta tcccgtgttg acgccgggca agagcaactc ggtcgccgca
     2161 tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg
     2221 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
     2281 ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca
     2341 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
     2401 acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa
     2461 ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg gaggcggata
     2521 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
     2581 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc
     2641 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
     2701 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt
     2761 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     2821 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     2881 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     2941 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     3001 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     3061 tccttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     3121 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     3181 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     3241 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     3301 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     3361 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     3421 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     3481 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     3541 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
     3601 gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg
     3661 agtcagtgag cgaggaagcg gaagagcgcc tgatgcggta ttttctcctt acgcatctgt
     3721 gcggtatttc acaccgcata tggtgcactc tcagtacaat ctgctctgat gccgcatagt
     3781 taagccagta tacactccgc tatcgctacg tgactgggtc atggctgcgc cccgacaccc
     3841 gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca
     3901 agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg
     3961 cgcgaggcag cagatcaatt cgcgcgcgaa ggcgaagcgg catgcataat gtgcctgtca
     4021 aatggacgaa gcagggattc tgcaaaccct atgctactcc gtcaagccgt caattgtctg
     4081 attcgttacc aattatgaca acttgacggc tacatcattc actttttctt cacaaccggc
     4141 acggaactcg ctcgggctgg ccccggtgca ttttttaaat acccgcgaga aatagagttg
     4201 atcgtcaaaa ccaacattgc gaccgacggt ggcgataggc atccgggtgg tgctcaaaag
     4261 cagcttcgcc tggctgatac gttggtcctc gcgccagctt aagacgctaa tccctaactg
     4321 ctggcggaaa agatgtgaca gacgcgacgg cgacaagcaa acatgctgtg cgacgctggc
     4381 gatatcaaaa ttgctgtctg ccaggtgatc gctgatgtac tgacaagcct cgcgtacccg
     4441 attatccatc ggtggatgga gcgactcgtt aatcgcttcc atgcgccgca gtaacaattg
     4501 ctcaagcaga tttatcgcca gcagctccga atagcgccct tccccttgcc cggcgttaat
     4561 gatttgccca aacaggtcgc tgaaatgcgg ctggtgcgct tcatccgggc gaaagaaccc
     4621 cgtattggca aatattgacg gccagttaag ccattcatgc cagtaggcgc gcggacgaaa
     4681 gtaaacccac tggtgatacc attcgcgagc ctccggatga cgaccgtagt gatgaatctc
     4741 tcctggcggg aacagcaaaa tatcacccgg tcggcaaaca aattctcgtc cctgattttt
     4801 caccaccccc tgaccgcgaa tggtgagatt gagaatataa cctttcattc ccagcggtcg
     4861 gtcgataaaa aaatcgagat aaccgttggc ctcaatcggc gttaaacccg ccaccagatg
     4921 ggcattaaac gagtatcccg gcagcagggg atcattttgc gcttcagcca tacttttcat
     4981 actcccgcca ttcagag
//
