LOCUS       sequence-300638-        5423 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses MlaF(1-250)-GCN4.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5423)
  AUTHORS   Kolich LR, Chang YT, Coudray N, Giacometti SI, MacRae MR, Isom GL, 
            Teran EM, Bhabha G, Ekiert DC
  TITLE     Structure of MlaFB uncovers novel mechanisms of ABC transporter 
            regulation.
  JOURNAL   Elife. 2020 Jun 30;9. pii: 60030. doi: 10.7554/eLife.60030.
  PUBMED    32602838
REFERENCE   2  (bases 1 to 5423)
  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: "Elife."; 
            date: "2020-06-30"; volume: "9. pii: 60030. doi"; pages: " 
            10.7554/eLife.60030"
COMMENT     Sequence Label: pBEL1833
FEATURES             Location/Qualifiers
     source          1..5423
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        10..294
                     /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     217..236
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             1105..1161
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility 
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     primer_bind     complement(1291..1308)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1461..1547
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1639..1666
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        1685..1776
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1777..1845,1846..2637)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(1995..2014)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2679..3134
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2766..2785)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2976..2997
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     rep_origin      3245..3832
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3733..3752
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3986..4003
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     4169..4191
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4329..4348
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             complement(4528..5406)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
ORIGIN
        1 cattcagaga agaaaccaat tgtccatatt gcatcagaca ttgccgtcac tgcgtctttt
       61 actggctctt ctcgctaacc aaaccggtaa ccccgcttat taaaagcatt ctgtaacaaa
      121 gcgggaccaa agccatgaca aaaacgcgta acaaaagtgt ctataatcac ggcagaaaag
      181 tccacattga ttatttgcac ggcgtcacac tttgctatgc catagcattt ttatccataa
      241 gattagcgga tcctacctga cgctttttat cgcaactctc tactgtttct ccatacccgt
      301 ttttttgggc tagccgaagg gtgaattatg gagcagtctg tggcgaattt agtcgatatg
      361 cgcgatgtca gttttacgcg tggcaatcgc tgcatcttcg ataatatttc cctgaccgtg
      421 ccgcgaggga agatcacggc gatcatgggg ccatcgggca tcggtaaaac gacgctactc
      481 cgtctgattg gcgggcaaat cgcaccagat catggtgaga tccttttcga tggtgagaat
      541 attccggcga tgtctcgttc gcgcctgtat acagtgcgca aacggatgag catgttattt
      601 cagtccgggg cgttgttcac tgatatgaac gtatttgaca acgtcgccta tccactgcgc
      661 gaacataccc aacttcccgc gccattgttg catagtacgg tgatgatgaa gctggaggcc
      721 gtggggctgc gtggagcggc taaactaatg ccttctgaac tttccggtgg gatggcgcgg
      781 cgtgcagcgc tggcgcgtgc gattgcgctg gagccggatc tcatcatgtt tgatgaacct
      841 tttgttgggc aagatcccat caccatgggc gtactggtga agctgatttc tgagctgaac
      901 agcgcgctgg gcgtgacttg tgtggtggtt tctcacgatg tgccggaagt gttaagtatt
      961 gcggatcacg cctggatcct ggcggacaaa aaaattgtcg ctcatggcag tgcccaggcg
     1021 ttgcaggcga atcctgatcc gcgcgtacgt cagtttctgg acgggatagc tgacgggcgt
     1081 atgaagcaac ttgaggacaa ggttgaagag ttattaagca aaaattatca tcttgaaaat
     1141 gaagtcgcgc gtttaaagaa gttagttggc gagcgctaag gtaccaaatt ccagaaaaga
     1201 gacgctgaaa agcgtctttt ttcgttttgg tcctggtctg aggaatggtt gtaagcttgg
     1261 ctgttttggc ggatgagaga agattttcag cctgatacag attaaatcag aacgcagaag
     1321 cggtctgata aaacagaatt tgcctggcgg cagtagcgcg gtggtcccac ctgaccccat
     1381 gccgaactca gaagtgaaac gccgtagcgc cgatggtagt gtggggtctc cccatgcgag
     1441 agtagggaac tgccaggcat caaataaaac gaaaggctca gtcgaaagac tgggcctttc
     1501 gttttatctg ttgtttgtcg gtgaacgctc tcctgagtag gacaaatccg ccgggagcgg
     1561 atttgaacgt tgcgaagcaa cggcccggag ggtggcgggc aggacgcccg ccataaactg
     1621 ccaggcatca aattaagcag aaggccatcc tgacggatgg cctttttgcg tttctacaaa
     1681 ctcttttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     1741 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
     1801 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
     1861 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
     1921 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
     1981 acttttaaag ttctgctatg tggcgcggta ttatcccgtg ttgacgccgg gcaagagcaa
     2041 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
     2101 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     2161 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
     2221 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     2281 gaagccatac caaacgacga gcgtgacacc acgatgcctg cagcaatggc aacaacgttg
     2341 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     2401 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     2461 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg
     2521 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     2581 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
     2641 tcagaccaag tttactcata tatactttag attgatttac gcgccctgta gcggcgcatt
     2701 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc
     2761 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     2821 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     2881 caaaaaactt gatttgggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
     2941 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
     3001 aacactcaac cctatctcgg gctattcttt tgatttataa gggattttgc cgatttcggc
     3061 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt
     3121 aacgtttaca atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     3181 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     3241 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     3301 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     3361 gctcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca
     3421 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     3481 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     3541 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     3601 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     3661 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     3721 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     3781 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     3841 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
     3901 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc
     3961 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgcct
     4021 gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatag ggtcatggct
     4081 gcgccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca
     4141 tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg
     4201 tcatcaccga aacgcgcgag gcagaaggag atggcgccca acagtccccc ggccacgggg
     4261 cctgccacca tacccacgcc gaaacaagcg ctcatgagcc cgaagtggcg agcccgatct
     4321 tccccatcgg tgatgtcggc gatataggcg ccagcaaccg cacctgtggc gccggtgatg
     4381 ccggccacga tgcgtccggc gtagaggatc tgctcatgtt tgacagctta tcatcgatgc
     4441 ataatgtgcc tgtcaaatgg acgaagcagg gattctgcaa accctatgct actccgtcaa
     4501 gccgtcaatt gtctgattcg ttaccaatta tgacaacttg acggctacat cattcacttt
     4561 ttcttcacaa ccggcacgga actcgctcgg gctggccccg gtgcattttt taaatacccg
     4621 cgagaaatag agttgatcgt caaaaccaac attgcgaccg acggtggcga taggcatccg
     4681 ggtggtgctc aaaagcagct tcgcctggct gatacgttgg tcctcgcgcc agcttaagac
     4741 gctaatccct aactgctggc ggaaaagatg tgacagacgc gacggcgaca agcaaacatg
     4801 ctgtgcgacg ctggcgatat caaaattgct gtctgccagg tgatcgctga tgtactgaca
     4861 agcctcgcgt acccgattat ccatcggtgg atggagcgac tcgttaatcg cttccatgcg
     4921 ccgcagtaac aattgctcaa gcagatttat cgccagcagc tccgaatagc gcccttcccc
     4981 ttgcccggcg ttaatgattt gcccaaacag gtcgctgaaa tgcggctggt gcgcttcatc
     5041 cgggcgaaag aaccccgtat tggcaaatat tgacggccag ttaagccatt catgccagta
     5101 ggcgcgcgga cgaaagtaaa cccactggtg ataccattcg cgagcctccg gatgacgacc
     5161 gtagtgatga atctctcctg gcgggaacag caaaatatca cccggtcggc aaacaaattc
     5221 tcgtccctga tttttcacca ccccctgacc gcgaatggtg agattgagaa tataaccttt
     5281 cattcccagc ggtcggtcga taaaaaaatc gagataaccg ttggcctcaa tcggcgttaa
     5341 acccgccacc agatgggcat taaacgagta tcccggcagc aggggatcat tttgcgcttc
     5401 agccatactt ttcatactcc cgc
//
