LOCUS       sequence-368145-        4719 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  FOLD6 ""hyperfolder"" green fluorescent protein: bacterial 
            expression plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4719)
  AUTHORS   Campbell BC, Paez-Segala MG, Looger LL, Petsko GA, Liu CF
  TITLE     Chemically stable fluorescent proteins for advanced microscopy.
  JOURNAL   Nat Methods. 2022 Nov 7. pii: 10.1038/s41592-022-01660-7. doi: 
            10.1038/s41592-022-01660-7.
  PUBMED    36344833
REFERENCE   2  (bases 1 to 4719)
  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: "Nat 
            Methods. 2022 Nov 7. pii: 10.1038/s41592-022-01660-7. doi: 
            10.1038/s41592-022-01660-7."
COMMENT     Sequence Label: pBAD-FOLD6
FEATURES             Location/Qualifiers
     source          1..4719
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        29..313
                     /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     236..255
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             356..373
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             join(383..385,386..388,389..1099)
                     /codon_start=1
                     /product="bright green-yellow fluorescent protein derived 
                     from GFP (Lam et al., 2012)"
                     /label=Clover
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
                     KFISTTGKLPVPWPTLVTTLGYGLAVFARYPDHMKQHDFFKSAMPEGYVQERTISFEDD
                     GYYKTRAEVKFEGDTLVNRIVLKGIDFKEDGNILGHKLEYNFNPHNVYITADKQKNGIK
                     ANFKTRHNVEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSHQSVLSKDPNEKRDHMVLL
                     EFVTAAGITHDMNELYK"
     primer_bind     complement(428..449)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     1036..1057
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1148..1165)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1318..1404
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1496..1523
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        1542..1633
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1634..1702,1703..2494)
                     /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(1852..1871)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2665..3253
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3154..3173
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3407..3424
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3439..3579
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3505..3524)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3665..3687
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(join(3843..4719,1..2))
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
ORIGIN
        1 atacttttca tactcccgcc attcagagaa gaaaccaatt gtccatattg catcagacat
       61 tgccgtcact gcgtctttta ctggctcttc tcgctaacca aaccggtaac cccgcttatt
      121 aaaagcattc tgtaacaaag cgggaccaaa gccatgacaa aaacgcgtaa caaaagtgtc
      181 tataatcacg gcagaaaagt ccacattgat tatttgcacg gcgtcacact ttgctatgcc
      241 atagcatttt tatccataag attagcggat cctacctgac gctttttatc gcaactctct
      301 actgtttctc catacccgtt ttttgggcta acaggaggaa ttaaccatgg ggagccacca
      361 tcaccatcac catggcagat ctatggtgag caagggcgag gagctgttca ccggggtggt
      421 gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg tccgcggcga
      481 gggcgagggc gatgccacca acggcaagct gaccctgaag ttcatctcca ccaccggcaa
      541 gctgcccgtg ccctggccca ccctcgtgac caccttaggc tacggcctgg ccgtgttcgc
      601 ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta
      661 cgtccaggag cgcaccatct ctttcgagga cgacggttac tacaagaccc gcgccgaggt
      721 gaagttcgag ggcgacaccc tggtgaaccg catcgtgctg aagggcatcg acttcaagga
      781 ggacggcaac atcctggggc acaagctgga gtacaacttc aacccccaca acgtctatat
      841 cacggccgac aagcagaaga acggcatcaa ggctaacttc aagacccgcc acaacgttga
      901 ggacggcggc gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc
      961 cgtgctgctg cccgacaacc actacctgag ccatcagtcc gtcctgagca aagaccccaa
     1021 cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga ttacacatga
     1081 catgaacgag ctgtacaagt aagaattcga agcttggctg ttttggtgga tgagagaaga
     1141 ttttcagcct gatacagatt aaatcagaac gcagaagcgg tctgataaaa cagaatttgc
     1201 ctggcggcag tagcgcggtg gtcccacctg accccatgcc gaactcagaa gtgaaacgcc
     1261 gtagcgccga tggtagtgtg gggtctcccc atgcgagagt agggaactgc caggcatcaa
     1321 ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt ttatctgttg tttgtcggtg
     1381 aacgctctcc tgagtaggac aaatccgccg ggagcggatt tgaacgttgc gaagcaacgg
     1441 cccggagggt ggcgggcagg acgcccgcca taaactgcca ggcatcaaat taagcagaag
     1501 gccatcctga cggatggcct ttttgcgttt ctacaaactc ttttgtttat ttttctaaat
     1561 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     1621 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
     1681 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
     1741 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
     1801 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg
     1861 cgcggtatta tcccgtgttg acgccgggca agagcaactc ggtcgccgca tacactattc
     1921 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
     1981 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     2041 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     2101 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
     2161 tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact
     2221 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
     2281 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     2341 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     2401 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     2461 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
     2521 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     2581 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     2641 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     2701 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     2761 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt
     2821 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     2881 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     2941 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     3001 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     3061 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     3121 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     3181 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     3241 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     3301 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
     3361 ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag
     3421 cgaggaagcg gaagagcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc
     3481 acaccgcata tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagta
     3541 tacactccgc tatcgctacg tgactgggtc atggctgcgc cccgacaccc gccaacaccc
     3601 gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc
     3661 gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgaggcag
     3721 cagatcaatt cgcgcgcgaa ggcgaagcgg catgcataat gtgcctgtca aatggacgaa
     3781 gcagggattc tgcaaaccct atgctactcc gtcaagccgt caattgtctg attcgttacc
     3841 aattatgaca acttgacggc tacatcattc actttttctt cacaaccggc acggaactcg
     3901 ctcgggctgg ccccggtgca ttttttaaat acccgcgaga aatagagttg atcgtcaaaa
     3961 ccaacattgc gaccgacggt ggcgataggc atccgggtgg tgctcaaaag cagcttcgcc
     4021 tggctgatac gttggtcctc gcgccagctt aagacgctaa tccctaactg ctggcggaaa
     4081 agatgtgaca gacgcgacgg cgacaagcaa acatgctgtg cgacgctggc gatatcaaaa
     4141 ttgctgtctg ccaggtgatc gctgatgtac tgacaagcct cgcgtacccg attatccatc
     4201 ggtggatgga gcgactcgtt aatcgcttcc atgcgccgca gtaacaattg ctcaagcaga
     4261 tttatcgcca gcagctccga atagcgccct tccccttgcc cggcgttaat gatttgccca
     4321 aacaggtcgc tgaaatgcgg ctggtgcgct tcatccgggc gaaagaaccc cgtattggca
     4381 aatattgacg gccagttaag ccattcatgc cagtaggcgc gcggacgaaa gtaaacccac
     4441 tggtgatacc attcgcgagc ctccggatga cgaccgtagt gatgaatctc tcctggcggg
     4501 aacagcaaaa tatcacccgg tcggcaaaca aattctcgtc cctgattttt caccaccccc
     4561 tgaccgcgaa tggtgagatt gagaatataa cctttcattc ccagcggtcg gtcgataaaa
     4621 aaatcgagat aaccgttggc ctcaatcggc gttaaacccg ccaccagatg ggcattaaac
     4681 gagtatcccg gcagcagggg atcattttgc gcttcagcc
//
