LOCUS       sequence-232052-        6477 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  precusor alkaline phosphatase, phoA, with C169S, C179S, C287S, 
            C337S, A369C and A427C.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6477)
  AUTHORS   Bariya P, Randall LL
  TITLE     Co-assembly of SecYEG and SecA fully restores the properties of the 
            native translocon.
  JOURNAL   J Bacteriol. 2018 Oct 1. pii: JB.00493-18. doi: 10.1128/JB.00493-18.
  PUBMED    30275279
REFERENCE   2  (bases 1 to 6477)
  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: "J 
            Bacteriol. 2018 Oct 1. pii: JB.00493-18. doi: 10.1128/JB.00493-18."
COMMENT     Sequence Label: pAL928
FEATURES             Location/Qualifiers
     source          1..6477
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     8..27
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     103..122
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        103..121
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     sig_peptide     148..210
                     /codon_start=1
                     /product="signal sequence of E. coli alkaline phosphatase 
                     (Inouye et al., 1982)"
                     /label=phoA signal sequence
                     /translation="MKQSTIALALLPLLFTPVTKA"
     primer_bind     complement(2598..2616)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2684..2788
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2789..2857,2858..3649)
                     /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(3007..3026)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3820..4408
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4309..4328
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4562..4579
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4594..4734
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4660..4679)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4820..4842
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4836..5027)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 ccccatcggt gatgtcggcg atataggcgc cagcaaccgc acctgtggcg ccggtgatgc
       61 cggccacgat gcgtccggcg tagaggatcg atcccgcgaa attaatacga ctcactatag
      121 ggagaccacg tacatggaga aaataaagtg aaacaaagca ctattgcact ggcactctta
      181 ccgttactgt ttacccctgt gacaaaagcc cggacaccag aaatgcctgt tctggaaaac
      241 cgggctgctc agggcgatat tactgcaccc ggcggtgctc gccgtttaac gggtgatcag
      301 actgccgctc tgcgtgattc tcttagcgat aaacctgcaa aaaatattat tttgctgatt
      361 ggcgatggga tgggggactc ggaaattact gccgcacgta attatgccga aggtgcgggc
      421 ggctttttta aaggtataga tgccttaccg cttaccgggc aatacactca ctatgcgctg
      481 aataaaaaaa ccggcaaacc ggactacgtc accgactcgg ctgcatcagc aaccgcctgg
      541 tcaaccggtg tcaaaaccta taacggcgcg ctgggcgtcg atattcacga aaaagatcac
      601 ccaacgattc tggaaatggc aaaagccgca ggtctggcga ccggtaacgt ttctaccgca
      661 gagttgcagg atgccacgcc cgctgcgctg gtggcacatg tgacctcgcg caaatcctac
      721 ggtccgagcg cgaccagtga aaaatctccg ggtaacgctc tggaaaaagg cggaaaagga
      781 tcgattaccg aacagctgct taacgctcgt gccgacgtta cgcttggcgg cggcgcaaaa
      841 acatttgctg aaacggcaac cgctggtgaa tggcagggaa aaacgctgcg tgaacaggca
      901 caggcgcgtg gttatcagtt ggtgagcgat gctgcctcac tgaattcggt gacggaagcg
      961 aatcagcaaa aacccctgct tggcctgttt gctgacggca atatgccagt gcgctggcta
     1021 ggaccgaaag caacgtacca tggcaatatc gataagcccg cagtcacctc tacgccaaat
     1081 ccgcaacgta atgacagtgt accaaccctg gcgcagatga ccgacaaagc cattgaattg
     1141 ttgagtaaaa atgagaaagg ctttttcctg caagttgaag gtgcgtcaat cgataaacag
     1201 gatcatgctg cgaatccttc tgggcaaatt ggcgagacgg tcgatctcga tgaagccgta
     1261 caacgggcgc tggaattcgc taaaaaggag ggtaacacgc tggtcatagt cacctgtgat
     1321 cacgcccacg ccagccagat tgttgcgccg gataccaaag ctccgggcct cacccaggcg
     1381 ctaaatacca aagatggcgc agtgatggtg atgagttacg ggaactccga agaggattca
     1441 caagaacata ccggcagtca gttgcgtatt gcggcgtatg gcccgcattg tgccaatgtt
     1501 gttggactga ccgaccagac cgatctcttc tacaccatga aagccgctct ggggctgaaa
     1561 taaaaccgcg cccggcagtg aattttcgct gccgggtggt ttttttgctg ttagcaacca
     1621 gacttaatgg cagatcacgg gcgcatacgc tcatggttaa aacatgaaga gggatggtgc
     1681 tatgaaaata acattactgg ttaccttgct tttcggtctg gtttttttaa ccaccgtcgg
     1741 cgctgccgag agaactttaa ccccacaaca acagcgtatg acctcctgta atcagcaggc
     1801 gacggcgcag gcgttgaaag gggatgctcg taagacctac atgagtgatt gcctgaagaa
     1861 cagcaagtct gcgcctggcg aaaaaagttt gacgccacag cagcaaaaga tgcgcgaatg
     1921 caataatcaa gcaacacaac aatctctgaa aggtgatgat cgtaataagt ttatgagtgc
     1981 ctgcctcaag aaagccgcct gatacctgat agtgctaacg ggtgagctac gaaaatggct
     2041 cacccgaaat atcatacttc tgcctttagc tccgtctcta taatttggga aaattgtttc
     2101 tgaatgttcc caaaaataat gaatgatgaa aactttttca aaaaagcggc ggcgcacggg
     2161 gaggaacctc ctttaactcc tcaaaacgaa catcagcggt ccgggctgcg cttcgcccgt
     2221 cgcgtcagac taccccgtgc ggttggcctg gctggcatgt tcttaccgat tgcttcaacg
     2281 ctggtttcac acccgccgcc gggctggtgg tggctggtgt tggtcggctg ggcgttcgtc
     2341 tggccgcatt tagcctggca gatagcgagc agggccgtcg atccgcttag ccgggaaatt
     2401 tacaacttaa aaaccgatgc agtattagcg ggaatgtggg taggcgtaat gggcgtaaac
     2461 gtgctgcctt ccaccgcgat gttgatgatt atgtgtctga atttgatggg ggcaggcggc
     2521 ccccgtctgt ttgtcgcggg tctggtgttg atggtggttt cctgccttgt caccctcgaa
     2581 ttcttgaaga cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat
     2641 aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     2701 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     2761 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
     2821 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
     2881 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
     2941 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
     3001 agttctgcta tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc aactcggtcg
     3061 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
     3121 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
     3181 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
     3241 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
     3301 accaaacgac gagcgtgaca ccacgatgcc tgcagcaatg gcaacaacgt tgcgcaaact
     3361 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
     3421 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
     3481 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
     3541 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     3601 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
     3661 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     3721 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     3781 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     3841 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     3901 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     3961 tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     4021 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     4081 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     4141 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     4201 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     4261 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     4321 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     4381 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     4441 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     4501 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
     4561 cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg cggtattttc tccttacgca
     4621 tctgtgcggt atttcacacc gcatatggtg cactctcagt acaatctgct ctgatgccgc
     4681 atagttaagc cagtatacac tccgctatcg ctacgtgact gggtcatggc tgcgccccga
     4741 cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac
     4801 agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg
     4861 aaacgcgcga ggcagctgcg gtaaagctca tcagcgtggt cgtgaagcga ttcacagatg
     4921 tctgcctgtt catccgcgtc cagctcgttg agtttctcca gaagcgttaa tgtctggctt
     4981 ctgataaagc gggccatgtt aagggcggtt ttttcctgtt tggtcactga tgcctccgtg
     5041 taagggggat ttctgttcat gggggtaatg ataccgatga aacgagagag gatgctcacg
     5101 atacgggtta ctgatgatga acatgcccgg ttactggaac gttgtgaggg taaacaactg
     5161 gcggtatgga tgcggcggga ccagagaaaa atcactcagg gtcaatgcca gcgcttcgtt
     5221 aatacagatg taggtgttcc acagggtagc cagcagcatc ctgcgatgca gatccggaac
     5281 ataatggtgc agggcgctga cttccgcgtt tccagacttt acgaaacacg gaaaccgaag
     5341 accattcatg ttgttgctca ggtcgcagac gttttgcagc agcagtcgct tcacgttcgc
     5401 tcgcgtatcg gtgattcatt ctgctaacca gtaaggcaac cccgccagcc tagccgggtc
     5461 ctcaacgaca ggagcacgat catgcgcacc cgtggccagg acccaacgct gcccgagatg
     5521 cgccgcgtgc ggctgctgga gatggcggac gcgatggata tgttctgcca agggttggtt
     5581 tgcgcattca cagttctccg caagaattga ttggctccaa ttcttggagt ggtgaatccg
     5641 ttagcgaggt gccgccggct tccattcagg tcgaggtggc ccggctccat gcaccgcgac
     5701 gcaacgcggg gaggcagaca aggtataggg cggcgcctac aatccatgcc aacccgttcc
     5761 atgtgctcgc cgaggcggca taaatcgccg tgacgatcag cggtccaatg atcgaagtta
     5821 ggctggtaag agccgcgagc gatccttgaa gctgtccctg atggtcgtca tctacctgcc
     5881 tggacagcat ggcctgcaac gcgggcatcc cgatgccgcc ggaagcgaga agaatcataa
     5941 tggggaaggc catccagcct cgcgtcgcga acgccagcaa gacgtagccc agcgcgtcgg
     6001 ccgccatgcc ggcgataatg gcctgcttct cgccgaaacg tttggtggcg ggaccagtga
     6061 cgaaggcttg agcgagggcg tgcaagattc cgaataccgc aagcgacagg ccgatcatcg
     6121 tcgcgctcca gcgaaagcgg tcctagccga aaatgaccca gagcgctgcc ggcacctgtc
     6181 ctacgagttg catgataaag aagacagtca taagtgcggc gacgatagtc atgccccgcg
     6241 cccaccggaa ggagctgact gggttgaagg ctctcaaggg catcggtcga cgctctccct
     6301 tatgcgactc ctgcattagg aagcagccca gtagtaggtt gaggccgttg agcaccgccg
     6361 ccgcaaggaa tggtgcatgc aaggagatgg cgcccaacag tcccccggcc acggggcctg
     6421 ccaccatacc cacgccgaaa caagcgctca tgagcccgaa gtggcgagcc cgatctt
//
