LOCUS       sequence-459798-        6985 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Vector for bacterial expression of a protein carrying the 
            non-canonical amino acid p-(phenylazo)-L-phenylalanine (encoding the
            OmpA signal sequence for periplasmic secretion).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6985)
  AUTHORS   Mayrhofer P, Anneser MR, Schira K, Sommer CA, Theobald I, Schlapschy
            M, Achatz S, Skerra A
  TITLE     Protein purification with light via a genetically encoded azobenzene
            side chain.
  JOURNAL   Nat Commun. 2024 Dec 18;15(1):10693. doi: 
            10.1038/s41467-024-55212-y.
  PUBMED    39695158
REFERENCE   2  (bases 1 to 6985)
  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 
            Commun."; date: "2024-12-18"; volume: "15(1):10693. doi"; pages: " 
            10.1038/s41467-024-55212-y"
COMMENT     Sequence Label: pSB19-PapRS#34-OmpA
FEATURES             Location/Qualifiers
     source          1..6985
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             144..1226
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=lacI
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(163..182)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     protein_bind    1239..1260
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1275..1280,1281..1298,1299..1305)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    1313..1329
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     1318..1340
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1337..1353
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1337..1353
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     sig_peptide     1398..1460
                     /codon_start=1
                     /product="signal peptide from the E. coli outer membrane 
                     protein OmpA"
                     /label=OmpA signal peptide
                     /translation="MKKTAIAIAVALAGFATVAQA"
     CDS             1522..1545
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     RBS             3059..3067
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     promoter        complement(3122..3406)
                     /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     complement(3180..3199)
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             3433..4311
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPSDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        4604..4708
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4709..4777,4778..5569)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(4927..4946)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     CDS             5594..6253
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHD*FRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(5660..5679)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      6379..6967
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6868..6887
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgacccgaca ccataacgct
       61 cggttgccgc ggtaccatgg cgcaaaacct ttcgcggtat ggcatgatag cgcccggaag
      121 agagtcaatt cagggtggtg aatgtgaaac cagtaacgtt atacgatgtc gcagagtatg
      181 ccggtgtctc ttatcagacc gtttcccgcg tggtgaacca ggccagccac gtttctgcga
      241 aaacgcggga aaaagtggaa gcggcgatgg cggagctgaa ttacattccc aaccgcgtgg
      301 cacaacaact ggcgggcaaa cagtcgttgc tgattggcgt tgccacctcc agtctggccc
      361 tgcacgcgcc gtcgcaaatt gtcgcggcga ttaaatctcg cgccgatcaa ctgggtgcca
      421 gcgtggtggt gtcgatggta gaacgaagcg gcgtcgaagc ctgtaaagcg gcggtgcaca
      481 atcttctcgc gcaacgcgtc agtgggctga tcattaacta tccgctggat gaccaggatg
      541 ccattgctgt ggaagctgcc tgcactaatg ttccggcgtt atttcttgat gtctctgacc
      601 agacacccat caacagtatt attttctccc atgaagacgg tacgcgactg ggcgtggagc
      661 atctggtcgc attgggtcac cagcaaatcg cgctgttagc gggcccatta agttctgtct
      721 cggcgcgtct gcgtctggct ggctggcata aatatctcac tcgcaatcaa attcagccga
      781 tagcggaacg ggaaggcgac tggagtgcca tgtccggttt tcaacaaacc atgcaaatgc
      841 tgaatgaggg catcgttccc actgcgatgc tggttgccaa cgatcagatg gcgctgggcg
      901 caatgcgcgc cattaccgag tccgggctgc gcgttggtgc ggatatctcg gtagtgggat
      961 acgacgatac cgaagacagc tcatgttata tcccgccgtt aaccaccatc aaacaggatt
     1021 ttcgcctgct ggggcaaacc agcgtggacc gcttgctgca actctctcag ggccaggcgg
     1081 tgaagggcaa tcagctgttg cccgtctcac tggtgaaaag aaaaaccacc ctggcgccca
     1141 atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg
     1201 tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat
     1261 taggcacccc aggctttaca ctttatgctt ccggctcgta taatgtgtgg aattgtgagc
     1321 ggataacaat ttcacacagg aaacagctat gaccatgatt acggattcac tggaactcta
     1381 gataacgagg gcaaaaaatg aaaaagacag ctatcgcgat tgcagtggca ctggctggtt
     1441 tcgctaccgt agcgcaggcc tgagaccaga attcgagctc ggtacccggg gatccctcga
     1501 ggtcgacctg caggcagcgc ttggagtcat ccacaattcg agaaataata agcttgacct
     1561 gtgaagtgaa aaatggcgca cattgtgcga catttttttt gtctgccgtt taccgctact
     1621 gcgtcacgga tcgccactgc tagcagttag taaaaaaaat ccttagcttt cgctaaggat
     1681 tgagctcatt acaggttggt gctaatacca ttgtaatagc tctcgctacg tgctgcacgt
     1741 ttaatgtttt tgaagtcgtg tttaactttc agcaggcgtt cgagaccaaa acctgcacca
     1801 atccacggtt tatcaatacc ccattcgcga tccagaggaa tcggaccaac aactgcgcta
     1861 ctcagttcca gatcaccatg cataacatcc agggtatcac caaacaccat gcagctatca
     1921 ccaacaattt tgaaatcgat gcccagatgg ttcagaaaat cggtaataat gctttccaga
     1981 ttttcacgcg tacaaccgct acccatctgg ccaaaagcca gcatggtaaa ttcttccagg
     2041 tgttctttac catcactttc tttgcggtag cacggaccaa tttcaaagat tttaatcgga
     2101 tccggcaggg cacgatccag tttgcggcta tagttataca gactcggggt cagcatcgga
     2161 cgcagacaaa ggtttttatc cacgcgaaag atttgtttgc tcagttcggt atcgttatca
     2221 atgcccatac gttcaatata ttccagcgga atcagaatcg ggcttttgat ttccaggaaa
     2281 ccacgatcca caaaaaaacg ggtgatttca cgttccagtt tacccagata gttttcacgt
     2341 tcttcggcat aaatctgttg caggtctttt ttacgacggc tcagcagttc gctttccagt
     2401 tcacgaaacg gtttaccgct attcaggcta atttcatctt tcggattcag cagaacttcc
     2461 aggcgatcgg tctgagaccg ggtcagtgcc ggtgcacttg cctgaaccgg tgcgctcatg
     2521 ctggtaatcg gattggtatt acctttaacc agtgcgcttg cggttgcacc ggttgaaatg
     2581 ctgctaatgc tggtgctcac gcttgccgga acgctaacgc tttcttgggt gctaaccgga
     2641 attgccggtg aaaatttgct accgctcggc tgtgcctgtg ctgcttcggt attttccagc
     2701 ggtttcggtg cacgggcaac gcttttcggc attgcttttt tggtacgtgt cggtgcgcta
     2761 acaactttaa ctttaacgct ggtctgatct tcattggctt tggtcagaaa tttgttcaga
     2821 tcctcatcac taacacggca acgtttacag gttttacgat atttgtgatg acgcagtgca
     2881 cgtgcggtac gactgctacg gctattattc accaccagat gatcaccaca tgccatttca
     2941 atatagattt tgctgcggct cacttcatga tgtttgattt tatgaatggt gccggtacgg
     3001 ctcatccaca gaccggttgc gctaatcagg gtattcagcg gttttttatc catatcgtac
     3061 ctccttaaat ttctaggttg tgacctaggc taattcctcc tgttagccca aaaaaacggg
     3121 tatggagaaa cagtagagag ttgcgataaa aagcgtcagg taggatccgc taatcttatg
     3181 gataaaaatg ctatggcata gcaaagtgtg acgccgtgca aataatcaat gtggactttt
     3241 ctgccgtgat tatagacact tttgttacgc gtttttgtca tggctttggt cccgctttgt
     3301 tacagaatgc ttttaataag cggggttacc ggtttggtta gcgagaagag ccagtaaaag
     3361 acgcagtgac ggcaatgtct gatgcaatat ggacaattgg tttcttctct gaatggcggg
     3421 agtatgaaaa gtatggctga agcgcaaaat gatcccctgc tgccgggata ctcgtttaat
     3481 gcccatctgg tggcgggttt aacgccgatt gaggccaacg gttatctcga tttttttatc
     3541 gaccgaccgc tgggaatgaa aggttatatt ctcaatctca ccattcgcgg tcagggggtg
     3601 gtgaaaaatc agggacgaga atttgtttgc cgaccgagtg atattttgct gttcccgcca
     3661 ggagagattc atcactacgg tcgtcatccg gaggctcgcg aatggtatca ccagtgggtt
     3721 tactttcgtc cgcgcgccta ctggcatgaa tggcttaact ggccgtcaat atttgccaat
     3781 acggggttct ttcgcccgga tgaagcgcac cagccgcatt tcagcgacct gtttgggcaa
     3841 atcattaacg ccgggcaagg ggaagggcgc tattcggagc tgctggcgat aaatctgctt
     3901 gagcaattgt tactgcggcg catggaagcg attaacgagt cgctccatcc accgatggat
     3961 aatcgggtac gcgaggcttg tcagtacatc agcgatcacc tggcagacag caattttgat
     4021 atcgccagcg tcgcacagca tgtttgcttg tcgccgtcgc gtctgtcaca tcttttccgc
     4081 cagcagttag ggattagcgt cttaagctgg cgcgaggacc aacgtatcag ccaggcgaag
     4141 ctgcttttga gcaccacccg gatgcctatc gccaccgtcg gtcgcaatgt tggttttgac
     4201 gatcaactct atttctcgcg ggtatttaaa aaatgcaccg gggccagccc gagcgagttc
     4261 cgtgccggtt gtgaagaaaa agtgaatgat gtagccgtca agttgtcata attgggtacg
     4321 aagcctcgag tcggtgccga tttcggccta ttggttaaaa aatgagctga gttctagtaa
     4381 aaaaaatcct tagctttcgc taaggatctg cagtggcgga aaccccggga atctaacccg
     4441 gctgaacgga tttagagtcc attcgatcta catgatcagg tttccgaatt cagcgttaca
     4501 agtattacac aaagtttttt atgttgagaa tatttttttg atggggcatg gcgcaaaacc
     4561 tttcgcggta tggcatgcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     4621 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     4681 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
     4741 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
     4801 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
     4861 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
     4921 agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg
     4981 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
     5041 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
     5101 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
     5161 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
     5221 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact
     5281 attaactggc gaactactta ctctagcttc ccggcaacaa ttgatagact ggatggaggc
     5341 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
     5401 taaatctgga gccggtgagc gtggctctcg cggtatcatt gcagcactgg ggccagatgg
     5461 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     5521 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaaa gatctatatg
     5581 tcgactagag gatatggaga aaaaaatcac tggatatacc accgttgata tatcccaatg
     5641 gcatcgtaaa gaacattttg aggcatttca gtcagttgct caatgtacct ataaccagac
     5701 cgttcagctg gatattacgg cctttttaaa gaccgtaaag aaaaataagc acaagtttta
     5761 tccggccttt attcacattc ttgcccgcct gatgaatgct catccggagt tccgtatggc
     5821 aatgaaagac ggtgagctgg tgatatggga tagtgttcac ccttgttaca ccgttttcca
     5881 tgagcaaact gaaacgtttt catcgctctg gagtgaatac cacgactagt tccggcagtt
     5941 tctacacata tattcgcaag atgtggcgtg ttacggtgaa aacctggcct atttccctaa
     6001 agggtttatt gagaatatgt ttttcgtctc agccaatccc tgggtgagtt tcaccagttt
     6061 tgatttaaac gtggccaata tggacaactt cttcgccccc gttttcacta tgggcaaata
     6121 ttatacgcaa ggcgacaagg tgctgatgcc gctggcgatt caggttcatc atgccgtttg
     6181 tgatggcttc catgtcggca gaatgcttaa tgaattacaa cagtactgcg atgagtggca
     6241 gggcggggcg taatagcttc actagtttaa aaggatctag gtgaagatcc tttttgataa
     6301 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     6361 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     6421 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     6481 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     6541 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     6601 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     6661 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     6721 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     6781 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     6841 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     6901 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     6961 atggaaaaac gccagcaacg cggcc
//
