LOCUS       sequence-343843-        4531 bp    DNA     circular SYN 14-APR-2025
DEFINITION  modified pMD18-T containing pyrF and its native promoter, Ampr.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4531)
  AUTHORS   Lin L, Chen J, Mitra R, Gao Q, Cheng F, Xu T, Zuo Z, Xiang H, Han J
  TITLE     Optimising PHBV biopolymer production in haloarchaea via 
            CRISPRi-mediated redirection of carbon flux.
  JOURNAL   Commun Biol. 2021 Aug 25;4(1):1007. doi: 10.1038/s42003-021-02541-z.
  PUBMED    34433872
REFERENCE   2  (bases 1 to 4531)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Commun 
            Biol."; date: "2021-08-25"; volume: "4(1):1007. doi"; pages: " 
            10.1038/s42003-021-02541-z"
COMMENT     Sequence Label: pMD18-T-pyrF
FEATURES             Location/Qualifiers
     source          1..4531
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(1802..1818)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1802..1818)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1815..1837)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1826..1842
                     /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)."
     promoter        complement(join(1850..1856,1857..1874,1875..1880))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1895..1916
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2033..2050)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2204..2792)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2284..2303)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2963..3754,3755..3823))
                     /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     3586..3605
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3824..3928)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3996..4014
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4052..4074)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4174..4193
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4387..4409
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4401..4418
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4402..4418
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 gtacctcgta tgccgccttc tcgattatcg cggcgggggc aacctaccgt tccgggtgga
       61 ttagcgaccg actcggcgtc cgacggtggt tcctcgtctc gccgcttctc gtcgccgtgt
      121 cctacgccgt gctcccaatc ttcggtccac tggcgttccc ggtatttgcc gtcgttcgcg
      181 gcgtaaacaa cgtctcgggc gtcctcggaa accagtacat caacgaccgt gtcgactcgg
      241 ttgggcgcgc aaccgtcctc tccgccgccg ggatgctcta ctcgctcgcc gtcgttccgt
      301 tcgaactcgc gggtggtgtc gtcgccgacg tggtctctcc aaccggcgca ctcgccctat
      361 tcggcggggt tcttgcggtc ggtgcaggcg tgttgtgggt tgtcgaacgc ccaatctgag
      421 ttttcttgtg tgcggttccc ttgcgtacag ttcccagcga ccttcccggc tgcgacgaag
      481 ggaaaacccc ttacatacgg tcgtcgcact cgcgggcaat gggcttcttt gacgaccttc
      541 aggaccgcat cgagaaggtg gacagtgtcg tctcggtcgg actcgacgcc gacctcgacc
      601 ggattcccga acacctccgc gacgaggacc tgccgcggtg ggcgttcaac cgccgcatca
      661 tcgacgcgac gcacgaacac gcggcggcct acaagccgaa cgcggccttc tacgaggacg
      721 aagacggctg gcgctcgctc cgcgagacca tcgcctacgc acacggaaag gatgttcccg
      781 tcctcctcga cgccaagcgc gccgacatcg gcaacacgac gcgacagtac gcgaacctgc
      841 ttgacgacgc cgatgccatc accgtcaacc cctacatggg ccgcgattcc atccagccgt
      901 tcctcgcccg cgaggacaag ggcgtgttcg tcctctgtcg cacctctaac ccgggcagcg
      961 cggacctcca gtcgctcgaa ctcgcctccg gagaacccct gtacgagcgc gtcgcggcgc
     1021 tctgcgacct ctggaacgaa aacgacaacg tcggactcgt cgtcggcgcg accggccccg
     1081 aagagttgga agacctccgc gagcaggtcc ccgacctgcc gtttctcgtc cccggcgtcg
     1141 gcgcacaggg tggcgatgcc gaggcggcag tcgagtacgg actggcagac ggcgtcggtc
     1201 tcgtgaactc ctcacgcggc atcatcttcg ccggcgaaga ccagggagaa cacttcgaca
     1261 aggctgcggg acaggcggcg aagcgactca aagagcggct gaaccagtac cggtaagtct
     1321 gcggaaatca cagactggca ctggactgta ccgctctcgg cgggacgttt accagcgtgc
     1381 gtcaccaact acgtccgtgg accgagaccg acttatcatg ggcctcgcgg cggtgttcgc
     1441 gggtattacc atgctcctcg tcgtgttggc attcgcctac cgacagctgt tgttgttgtt
     1501 cgtagctgtg cccttcggcg cgacgacgta cttcatgtac tcggacgtga gcggccggtt
     1561 cgaagaacgg tttagacggc gagttcaggc ggatgctggc ggggtcggct ttcagtcggg
     1621 cggtggcggc ccacagcagc gcgaaaccgg cgggttcggt gcagggccac gacgtggtcc
     1681 ccgagaaggc cccggtgcag gacggttcgg gcgcgccgga cgacagggtg gaccgcaaag
     1741 tggcgcacgt actgaaatct ctagaggatc cccgggtacc gagctcgaat tcgtaatcat
     1801 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
     1861 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
     1921 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     1981 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
     2041 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     2101 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     2161 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     2221 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     2281 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     2341 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     2401 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     2461 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     2521 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     2581 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     2641 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     2701 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     2761 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     2821 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     2881 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     2941 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     3001 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     3061 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     3121 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     3181 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     3241 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     3301 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     3361 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     3421 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     3481 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     3541 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     3601 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     3661 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     3721 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     3781 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     3841 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     3901 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct
     3961 aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc
     4021 gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg
     4081 tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg
     4141 gtgttggcgg gtgtcggggc tggcttaact atgcggcatc agagcagatt gtactgagag
     4201 tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc
     4261 gccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc
     4321 tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg gtaacgccag
     4381 ggttttccca gtcacgacgt tgtaaaacga cggccagtgc caagcttgca tgcctgcagg
     4441 tcgacgattg cgtacatcct cgaccagccg atactccgtg ctgtcgcagt ggaccttggt
     4501 atccccgagt cagcgacaaa tacggccgtg a
//
