LOCUS       sequence-464727-        8530 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression plasmid of Cys-light LetA(Q180C,R380C) with LetB.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8530)
  AUTHORS   Santarossa CC, Li Y, Yousef S, Hasdemir HS, Rodriguez CC, Haase MB, 
            Baek M, Coudray N, Pavek JG, Focke KN, Silverberg AL, Bautista C, 
            Yeh J, Marty MT, Baker D, Tajkhorshid E, Ekiert DC, Bhabha G
  TITLE     LetA defines a structurally distinct transporter family involved in 
            lipid trafficking
  JOURNAL   bioRxiv 2025.03.21.644421
REFERENCE   2  (bases 1 to 8530)
  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: "bioRxiv 
            2025.03.21.644421"
COMMENT     Sequence Label: pBEL2820
FEATURES             Location/Qualifiers
     source          1..8530
                     /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             331..348
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             361..381
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(4398..4415)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      4568..4654
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      4746..4773
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        4792..4883
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4884..4952,4953..5744)
                     /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(5102..5121)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5786..6241
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5873..5892)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6083..6104
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     rep_origin      6352..6939
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6840..6859
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7093..7110
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     7276..7298
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7436..7455
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             complement(7635..8513)
                     /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 catcaccacc atcaccatca gcatcaacac
      361 gaaaatctgt attttcaggg gatggctctt aacacaccac aaattacgcc gacaaaaaag
      421 ataacagtga gggcaatcgg cgaggaactg ccgcgtggtg attaccaacg ttgcccgcaa
      481 tgtgacatgc tgtttagcct gcccgagata aattctcatc aaagtgccta ttgtccgcgc
      541 tgtcaggcaa aaattcgtga cgggcgcgac tggtcgctaa cgcgcctggc ggcaatggcc
      601 ttcaccatgc tgttgcttat gccgtttgcc tggggcgaac cgctgttgca tatctggctg
      661 ttaggcattc gtattgacgc caacgttatg caaggcatct ggcaaatgac caaacagggc
      721 gatgcgataa cggggtcgat ggtctttttc tctgttattg gtgccccgct cattctggtg
      781 acctccatag cttatttatg gtttggtaac cgactgggaa tgaatttacg tccggtactg
      841 ctgatgcttg agcgacttaa agagtgggta atgctcgata tctacctggt cggcattggc
      901 gttgcttcta taaaggtatg cgattatgcc catatccagg ctggtgtcgg cttgttctct
      961 tttgtggcgt tggtgatttt aacgacggtg acgttgtcac atcttaatgt cgaagaactg
     1021 tgggagcgat tttatccgca gcgccccgct tgacgaaggg tgaattatga cgcgtaggga
     1081 cgagaaactt cgtgtctgcc ttgggtgcca ttttaccggc tatccagatc agcgtggtcg
     1141 ctgcccgcgt tgccatatcc cgctacgcct gcgtcgccgt catagtctgc aaaaatcttg
     1201 ggcggcgctg ttagcgtcaa tcgttttatt gttacctgcc aacctgttgc ctatttctat
     1261 catttatctg aacggaggac ggcaggaaga tacaattctt tccggaatta tgtcgctggc
     1321 aagtagcaac attgcggttg caggaatcgt gtttatcgcc agtattctgg tcccgtttac
     1381 taaagtgatc gtcatgttca ctttactgtt gagcattcat tttaaatctc agcaaggttt
     1441 acgcacacgc attctgttac tgcggatggt gacctggatt ggtcgctggt cgatgctcga
     1501 cctgtttgtc atatctttaa ccatgtcgct gattaattgc gatcagatcc tcgcttttac
     1561 tatgggaccg gctgcgtttt atttcggcgc agcggtaatt ttgactattc ttgctgtgga
     1621 atggctggac agccgcttac tttgggatgc acatgagtca ggaaacgccc gcttcgacga
     1681 ctgaagcgca gattaaaaat aaacgccgta tctcaccttt ctggctgctg ccgttcatcg
     1741 cgctaatgat tgccagttgg ctgatttggg acagttatca ggaccggggt aataccgtca
     1801 ccatcgactt tatgtcggcg gatggtattg ttccaggccg tacgcctgtt cgttatcagg
     1861 gcgttgaagt cggaacagtg caggatatca gcctcagcga cgatcttcgt aagattgaag
     1921 tcaaggtcag catcaagtcc gatatgaaag atgcgctgcg cgaagagact cagttctggc
     1981 tggtgacgcc aaaagcatcg ttggcaggtg tctccgggct ggacgccctc gtcggtggga
     2041 actatatcgg catgatgccg ggtaaaggta aagagcagga tcactttgtc gcactcgata
     2101 cccaaccgaa atatcggctg gacaatggcg atctgatgat ccacctgcaa gcccccgatc
     2161 tcggttcgct gaacagcggt tcattggtct atttccgcaa gatcccggtg ggaaaagtct
     2221 acgactatgc catcaatccc aacaagcaag gcgtggtgat tgatgtcctg atcgagcggc
     2281 gttttaccga tctggtgaaa aaaggtagcc gtttctggaa cgtttccggc gttgatgcca
     2341 acgtcagtat cagtggcgcg aaggtgaaac tggaaagcct ggcggcactg gttaacggtg
     2401 cgattgcctt cgattcacca gaagagtcga aacctgccga ggcggaagat acctttggtc
     2461 tgtatgaaga tctggcccac agccagcgtg gcgtaataat aaaactggaa ctgccgagtg
     2521 gggccggatt aaccgccgac tcgacgccgt taatgtatca ggggctggaa gtcggacagc
     2581 tgactaaact ggatttaaat cctggtggta aagtcacagg ggaaatgacc gttgatccca
     2641 gcgtcgttac cctgcttcgt gaaaataccc gcatcgaatt acgcaacccg aaattatccc
     2701 ttagcgatgc taatctcagc gccctgctga ccggcaaaac cttcgagctg gtgcccggcg
     2761 atggcgagcc acgcaaagag ttcgttgttg tgccaggcga aaaagcactg ctgcatgaac
     2821 ctgatgttct gacgctgacc ctgaccgcgc cggaaagtta cggtattgat gcgggtcagc
     2881 cgctcattct tcacggcgtg caggtaggcc aggttatcga tcgtaaactc accagcaaag
     2941 gcgtcacctt taccgtcgcc atcgagcctc agcatcgcga actggtaaaa ggcgatagca
     3001 aatttgtcgt caacagccgt gtcgatgtga aggtggggct ggatggcgtt gagtttctcg
     3061 gtgccagcgc ctcagaatgg atcaatggcg ggatacgtat tctgccgggc gataaaggtg
     3121 agatgaaagc cagctatcca ctgtatgcca atttggaaaa agcgctggag aacagcctta
     3181 gcgatttacc caccacaacc gtgagtttga gtgcagagac gctgccggat gtgcaggcag
     3241 gatcggtagt gctgtaccgt aaatttgaag ttggtgaagt gattaccgtg cgtccgcgag
     3301 ctaacgcgtt tgatatcgat ctgcatatta agccggagta tcgcaacctt ctgaccagca
     3361 atagcgtgtt ctgggcagaa ggcggggcga aagttcagct gaatggtagt ggtctgaccg
     3421 tacaggcatc cccgctctcc agagcattaa agggagccat tagcttcgac aacctcagcg
     3481 gtgccagcgc cagtcagcgt aaaggcgata agcgtattct gtatgcttcc gaaacagcgg
     3541 cccgtgcggt tggcgggcag attacgcttc acgctttcga tgccggaaaa ctggcggtcg
     3601 ggatgccaat tcgctatctc ggtattgata tcgggcaaat ccagacgctg gatctgatta
     3661 ccgcacgcaa tgaagtacag gcaaaggcgg tactttaccc ggaatatgtc cagacctttg
     3721 ctcgcggtgg tacgcgcttc tcagtggtca caccgcaaat ttcggcagct ggcgttgagc
     3781 atcttgatac tatcctccag ccgtatatca acgtcgaacc aggccggggc aatcctcgcc
     3841 gcgactttga attacaagag gccaccatta ctgattcgcg ttacctggat ggcttaagca
     3901 ttattgttga agcgccggaa gccggttcgt taggcatcgg tacgcctgtg ctgttccgtg
     3961 gtctggaagt cggtacggtt acaggaatga cgctggggac attgtcagat cgcgtgatga
     4021 ttgcgatgcg catcagtaaa cgctatcaac acctggtgcg taacaattcc gtcttctggt
     4081 tggcatcggg ttacagtctg gactttggtc tgacgggcgg cgtagtgaaa accggcacct
     4141 ttaaccagtt tatccgtggc ggcatcgcct tcgccacgcc tccgggtacg ccactggcac
     4201 cgaaagccca ggaaggcaaa cacttcctgt tgcaggaaag tgaaccgaaa gagtggcgtg
     4261 aatggggaac tgcgcttccc aaataaggta ccaaattcca gaaaagagac gctgaaaagc
     4321 gtcttttttc gttttggtcc tggtctgagg aatggttgta agcttggctg ttttggcgga
     4381 tgagagaaga ttttcagcct gatacagatt aaatcagaac gcagaagcgg tctgataaaa
     4441 cagaatttgc ctggcggcag tagcgcggtg gtcccacctg accccatgcc gaactcagaa
     4501 gtgaaacgcc gtagcgccga tggtagtgtg gggtctcccc atgcgagagt agggaactgc
     4561 caggcatcaa ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt ttatctgttg
     4621 tttgtcggtg aacgctctcc tgagtaggac aaatccgccg ggagcggatt tgaacgttgc
     4681 gaagcaacgg cccggagggt ggcgggcagg acgcccgcca taaactgcca ggcatcaaat
     4741 taagcagaag gccatcctga cggatggcct ttttgcgttt ctacaaactc ttttgtttat
     4801 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     4861 aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct
     4921 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
     4981 atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta
     5041 agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc
     5101 tgctatgtgg cgcggtatta tcccgtgttg acgccgggca agagcaactc ggtcgccgca
     5161 tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg
     5221 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
     5281 ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca
     5341 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
     5401 acgacgagcg tgacaccacg atgcctgcag caatggcaac aacgttgcgc aaactattaa
     5461 ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg gaggcggata
     5521 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
     5581 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc
     5641 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
     5701 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt
     5761 actcatatat actttagatt gatttacgcg ccctgtagcg gcgcattaag cgcggcgggt
     5821 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     5881 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     5941 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     6001 ttgggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     6061 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
     6121 atctcgggct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
     6181 aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac gtttacaatt
     6241 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     6301 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     6361 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     6421 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct cagcagagcg
     6481 cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct
     6541 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     6601 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     6661 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     6721 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     6781 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     6841 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     6901 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     6961 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     7021 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
     7081 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcctgat gcggtatttt
     7141 ctccttacgc atctgtgcgg tatttcacac cgcatagggt catggctgcg ccccgacacc
     7201 cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac
     7261 aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac
     7321 gcgcgaggca gaaggagatg gcgcccaaca gtcccccggc cacggggcct gccaccatac
     7381 ccacgccgaa acaagcgctc atgagcccga agtggcgagc ccgatcttcc ccatcggtga
     7441 tgtcggcgat ataggcgcca gcaaccgcac ctgtggcgcc ggtgatgccg gccacgatgc
     7501 gtccggcgta gaggatctgc tcatgtttga cagcttatca tcgatgcata atgtgcctgt
     7561 caaatggacg aagcagggat tctgcaaacc ctatgctact ccgtcaagcc gtcaattgtc
     7621 tgattcgtta ccaattatga caacttgacg gctacatcat tcactttttc ttcacaaccg
     7681 gcacggaact cgctcgggct ggccccggtg cattttttaa atacccgcga gaaatagagt
     7741 tgatcgtcaa aaccaacatt gcgaccgacg gtggcgatag gcatccgggt ggtgctcaaa
     7801 agcagcttcg cctggctgat acgttggtcc tcgcgccagc ttaagacgct aatccctaac
     7861 tgctggcgga aaagatgtga cagacgcgac ggcgacaagc aaacatgctg tgcgacgctg
     7921 gcgatatcaa aattgctgtc tgccaggtga tcgctgatgt actgacaagc ctcgcgtacc
     7981 cgattatcca tcggtggatg gagcgactcg ttaatcgctt ccatgcgccg cagtaacaat
     8041 tgctcaagca gatttatcgc cagcagctcc gaatagcgcc cttccccttg cccggcgtta
     8101 atgatttgcc caaacaggtc gctgaaatgc ggctggtgcg cttcatccgg gcgaaagaac
     8161 cccgtattgg caaatattga cggccagtta agccattcat gccagtaggc gcgcggacga
     8221 aagtaaaccc actggtgata ccattcgcga gcctccggat gacgaccgta gtgatgaatc
     8281 tctcctggcg ggaacagcaa aatatcaccc ggtcggcaaa caaattctcg tccctgattt
     8341 ttcaccaccc cctgaccgcg aatggtgaga ttgagaatat aacctttcat tcccagcggt
     8401 cggtcgataa aaaaatcgag ataaccgttg gcctcaatcg gcgttaaacc cgccaccaga
     8461 tgggcattaa acgagtatcc cggcagcagg ggatcatttt gcgcttcagc catacttttc
     8521 atactcccgc
//
