LOCUS       sequence-235836-        4459 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Syb37 in e. coli cells, with Arabinose induction and 
            Ampicillin selection.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4459)
  AUTHORS   Brauer P, Parker JL, Gerondopoulos A, Zimmermann I, Seeger MA, Barr 
            FA, Newstead S
  TITLE     Structural basis for pH-dependent retrieval of ER proteins from the 
            Golgi by the KDEL receptor.
  JOURNAL   Science. 2019 Mar 8;363(6431):1103-1107. doi: 
            10.1126/science.aaw2859.
  PUBMED    30846601
REFERENCE   2  (bases 1 to 4459)
  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"; doi: "10.1126/science"; 
            journalName: "Science"; date: "2019-03-8- 8"; volume: "363"; issue: 
            "6431"; pages: "1103-1107"
COMMENT     Sequence Label: pBXPC3H-Syb37
FEATURES             Location/Qualifiers
     source          1..4459
                     /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"
     sig_peptide     319..384
                     /codon_start=1
                     /gene="pelB (fragment)"
                     /product="leader peptide for secretion"
                     /label=pelB signal sequence
                     /translation="MKYLLPTAAAGLLLLAAQPAMA"
     CDS             751..774
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             775..804
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     primer_bind     complement(860..877)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1030..1116
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1208..1235
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        1254..1345
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1346..1414,1415..2206)
                     /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(1564..1583)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2377..2965
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2866..2885
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3119..3136
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3151..3291
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3217..3236)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3377..3399
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3555..4433)
                     /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 tactggcact
       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 gaaatacctg ctgccgaccg ctgctgccgg actgctgctc
      361 ctcgctgccc agcctgcaat ggccagtcag gttcagctgg ttgagagcgg tggtggcctg
      421 gtccaagctg gcggttcgct gcgtctgagc tgcgccgcaa gcggtttccc ggtgaaacgt
      481 tggtctatga cttggtatcg tcaggcaccg ggcaaagaac gtgagtgggt cgcggcgatt
      541 cgtagcgctg gtcattggac gcattacgca gattctgtta agggccgctt taccatcagc
      601 cgcgacaacg cgaagaatac ggtctatttg cagatgaata gcctgaaacc ggaagatacc
      661 gcggtttact actgtaacgt gaaagacgaa ggtgacttct cttactggta cgactattgg
      721 ggccaaggta cccaagtgac tgtgagcgca ttagaagttt tgtttcaagg tccacatcat
      781 catcatcatc atcatcatca tcattgagtt taaacggtct ccagcttggc tgttttggcg
      841 gatgagagaa gattttcagc ctgatacaga ttaaatcaga acgcagaagc ggtctgataa
      901 aacagaattt gcctggcggc agtagcgcgg tggtcccacc tgaccccatg ccgaactcag
      961 aagtgaaacg ccgtagcgcc gatggtagtg tggggtctcc ccatgcgaga gtagggaact
     1021 gccaggcatc aaataaaacg aaaggctcag tcgaaagact gggcctttcg ttttatctgt
     1081 tgtttgtcgg tgaacgctct cctgagtagg acaaatccgc cgggagcgga tttgaacgtt
     1141 gcgaagcaac ggcccggagg gtggcgggca ggacgcccgc cataaactgc caggcatcaa
     1201 attaagcaga aggccatcct gacggatggc ctttttgcgt ttctacaaac tcttttgttt
     1261 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     1321 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
     1381 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     1441 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
     1501 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt
     1561 tctgctatgt ggcgcggtat tatcccgtgt tgacgccggg caagagcaac tcggtcgccg
     1621 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
     1681 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     1741 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
     1801 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     1861 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt
     1921 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
     1981 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     2041 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa
     2101 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     2161 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt
     2221 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     2281 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     2341 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     2401 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     2461 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     2521 tgtccttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     2581 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     2641 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     2701 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     2761 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     2821 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     2881 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     2941 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     3001 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa
     3061 ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag
     3121 cgagtcagtg agcgaggaag cggaagtgcg cctgatgcgg tattttctcc ttacgcatct
     3181 gtgcggtatt tcacaccgca tatggtgcac tctcagtaca atctgctctg atgccgcata
     3241 gttaagccag tatacactcc gctatcgcta cgtgactggg tcatggctgc gccccgacac
     3301 ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga
     3361 caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa
     3421 cgcgcgaggc agcagatcaa ttcgcgcgcg aaggcgaagc ggcatgcata atgtgcctgt
     3481 caaatggacg aagcagggat tctgcaaacc ctatgctact ccgtcaagcc gtcaattgtc
     3541 tgattcgtta ccaattatga caacttgacg gctacatcat tcactttttc ttcacaaccg
     3601 gcacggaact cgctcgggct ggccccggtg cattttttaa atacccgcga gaaatagagt
     3661 tgatcgtcaa aaccaacatt gcgaccgacg gtggcgatag gcatccgggt ggtgctcaaa
     3721 agcagcttcg cctggctgat acgttggtcc tcgcgccagc ttaagacgct aatccctaac
     3781 tgctggcgga aaagatgtga cagacgcgac ggcgacaagc aaacatgctg tgcgacgctg
     3841 gcgatatcaa aattgctgtc tgccaggtga tcgctgatgt actgacaagc ctcgcgtacc
     3901 cgattatcca tcggtggatg gagcgactcg ttaatcgctt ccatgcgccg cagtaacaat
     3961 tgctcaagca gatttatcgc cagcagctcc gaatagcgcc cttccccttg cccggcgtta
     4021 atgatttgcc caaacaggtc gctgaaatgc ggctggtgcg cttcatccgg gcgaaagaac
     4081 cccgtattgg caaatattga cggccagtta agccattcat gccagtaggc gcgcggacga
     4141 aagtaaaccc actggtgata ccattcgcga gcctccggat gacgaccgta gtgatgaatc
     4201 tctcctggcg ggaacagcaa aatatcaccc ggtcggcaaa caaattctcg tccctgattt
     4261 ttcaccaccc cctgaccgcg aatggtgaga ttgagaatat aacctttcat tcccagcggt
     4321 cggtcgataa aaaaatcgag ataaccgttg gcctcaatcg gcgttaaacc cgccaccaga
     4381 tgggcattaa acgagtatcc cggcagcagg ggatcatttt gcgcttcagc catacttttc
     4441 atactcccgc cattcagag
//
