LOCUS       sequence-251561-        4705 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4705)
  AUTHORS   Zuniga JE, Schmidt JJ, Fenn T, Burnett JC, Arac D, Gussio R, 
            Stafford RG, Badie SS, Bavari S, Brunger AT
  TITLE     A potent peptidomimetic inhibitor of botulinum neurotoxin serotype A
            has a very different conformation than SNAP-25 substrate.
  JOURNAL   Structure. 2008 Oct 8. 16(10):1588-97.
  PUBMED    18940613
REFERENCE   2  (bases 1 to 4705)
  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: "Structure. 
            2008 Oct 8. 16(10):1588-97."
COMMENT     Sequence Label: BoNT/A-LC
FEATURES             Location/Qualifiers
     source          1..4705
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(1..589)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(81..100)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(760..1551,1552..1620))
                     /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     1383..1402
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1621..1725)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1793..1811
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(1835..1849,1850..1855,1856..1872,1873..1879)
                     /label=T5 promoter
                     /note="bacteriophage T5 promoter for E. coli RNA 
                     polymerase, with embedded lac operator"
     protein_bind    1855..1871
                     /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)."
     protein_bind    1887..1903
                     /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)."
     CDS             3203..3220
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             3224..3241
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      3277..3371
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             3415..4074
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(3481..3500)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      4139..4225
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(4272..4294)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    4380..4520
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     4435..4454
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4535..4552)
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
       61 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
      121 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
      181 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
      241 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
      301 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
      361 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
      421 taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac
      481 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
      541 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
      601 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
      661 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
      721 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
      781 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
      841 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
      901 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
      961 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     1021 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     1081 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     1141 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     1201 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     1261 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     1321 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     1381 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     1441 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     1501 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     1561 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     1621 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     1681 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     1741 agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg
     1801 tatcacgagg cccttttgtc ttcacctcga gaaatcataa aaaatttatt tgctttgtga
     1861 gcggataaca attataatag attcaattgt gagcggataa caatttcaca cagaattcgc
     1921 aattaaggag ataataggta tgccatttgt taataaacaa tttaattata aagatcctgt
     1981 aaatggtgtt gatattgctt atataaaaat tccaaatgca ggacaaatgc aaccagtaaa
     2041 agcttttaaa attcataata aaatatgggt tattccagaa agagatacat ttacaaatcc
     2101 tgaagaagga gatttaaatc caccaccaga agcaaaacaa gttccagttt catattatga
     2161 ttcaacatat ttaagtacag ataatgaaaa agataattat ttaaagggag ttacaaaatt
     2221 atttgagaga atttattcaa ctgatcttgg aagaatgttg ttaacatcaa tagtaagggg
     2281 aataccattt tggggtggaa gtacaataga tacagaatta aaagttattg atactaattg
     2341 tattaatgtg atacaaccag atggtagtta tagatcagaa gaacttaatc tagtaataat
     2401 aggaccctca gctgatatta tacagtttga atgtaaaagc tttggacatg aagttttgaa
     2461 tcttacgcga aatggttatg gctctactca atacattaga tttagcccag attttacatt
     2521 tggttttgag gagtcacttg aagttgatac aaatcctctt ttaggtgcag gcaaatttgc
     2581 tacggatccg gcagtaactc tggcacacga actcatccac gctggtcacc gtctgtacgg
     2641 tatcgcgatt aatccaaata gggtttttaa agtaaatact aatgcctatt atgaaatgag
     2701 tgggttagaa gtaagctttg aggaacttag aacatttggg ggacatgatg caaagtttat
     2761 agatagttta caggaaaacg aatttcgtct atattattat aataagttta aagatatagc
     2821 aagtacactt aataaagcta aatcaatagt aggtactact gcttcattac agtatatgaa
     2881 aaatgttttt aaagagaaat atctcctatc tgaagataca tctggaaaat tttcggtaga
     2941 taaattaaaa tttgataagt tatacaaaat gttaacagag atttacacag aggataattt
     3001 tgttaagttt tttaaagtac ttaacagaaa aacatatttg aattttgata aagccgtatt
     3061 taagataaat atagtaccta aggtaaatta cacaatatat gatggattta atttaagaaa
     3121 tacaaattta gcagcaaact ttaatggtca aaatacagaa attaataata tgaattttac
     3181 taaactaaaa aattttactc ccctggttcc gcgtggatct gggcatcacc atcaccatca
     3241 ctaaggcctg cagccaagct taattagctg agcttggact cctgttgata gatccagtaa
     3301 tgacctcaga actccatctg gatttgttca gaacgctcgg ttgccgccgg gcgtttttta
     3361 ttggtgagaa tccaagctag cttggcgaga ttttcaggag ctaaggaagc taaaatggag
     3421 aaaaaaatca ctggatatac caccgttgat atatcccaat ggcatcgtaa agaacatttt
     3481 gaggcatttc agtcagttgc tcaatgtacc tataaccaga ccgttcagct ggatattacg
     3541 gcctttttaa agaccgtaaa gaaaaataag cacaagtttt atccggcctt tattcacatt
     3601 cttgcccgcc tgatgaatgc tcatccggaa tttcgtatgg caatgaaaga cggtgagctg
     3661 gtgatatggg atagtgttca cccttgttac accgttttcc atgagcaaac tgaaacgttt
     3721 tcatcgctct ggagtgaata ccacgacgat ttccggcagt ttctacacat atattcgcaa
     3781 gatgtggcgt gttacggtga aaacctggcc tatttcccta aagggtttat tgagaatatg
     3841 tttttcgtct cagccaatcc ctgggtgagt ttcaccagtt ttgatttaaa cgtggccaat
     3901 atggacaact tcttcgcccc cgttttcacc atgggcaaat attatacgca aggcgacaag
     3961 gtgctgatgc cgctggcgat tcaggttcat catgccgttt gtgatggctt ccatgtcggc
     4021 agaatgctta atgaattaca acagtactgc gatgagtggc agggcggggc gtaatttttt
     4081 taaggcagtt attggtgccc ttaaacgcct ggggtaatga ctctctagct tgaggcatca
     4141 aataaaacga aaggctcagt cgaaagactg ggcctttcgt tttatctgtt gtttgtcggt
     4201 gaacgctctc ctgagtagga caaatccgcc ctctagagct gcctcgcgcg tttcggtgat
     4261 gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg tctgtaagcg
     4321 gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc
     4381 gcagccatga cccagtcacg tagcgatagc ggagtgtata ctggcttaac tatgcggcat
     4441 cagagcagat tgtactgaga gtgcaccata tgcggtgtga aataccgcac agatgcgtaa
     4501 ggagaaaata ccgcatcagg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     4561 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     4621 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     4681 gtaaaaaggc cgcgttgctg gcgtt
//
