LOCUS       sequence-478138-        4107 bp    DNA     circular SYN 10-JUL-2025
DEFINITION  expresses N-His tagged NocM (acyl carrier protein) from Nodularia 
            sp. 06071.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4107)
  AUTHORS   Wang ML, Glasser NR, Nair MA, Krebs C, Martin Bollinger J Jr, 
            Balskus EP
  TITLE     Biochemical Studies of a Cyanobacterial Halogenase Support the 
            Involvement of a Dimetal Cofactor.
  JOURNAL   Biochemistry. 2025 May 20;64(10):2173-2180. doi: 
            10.1021/acs.biochem.4c00720. Epub 2025 Apr 29.
  PUBMED    40300765
REFERENCE   2  (bases 1 to 4107)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 10, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1021/acs.biochem.4c00720"; journalName: "Biochemistry"; date: 
            "2025-05-20- 20"; volume: "64"; issue: "10"; pages: "2173-2180"
COMMENT     Sequence Label: pCDFDuet-NHis6NocM-emptyMCS2
FEATURES             Location/Qualifiers
     source          1..4107
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        375..393
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    394..418
                     /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             527..571
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(609..627)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      623..670
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(841..1632)
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     promoter        complement(1633..1724)
                     /gene="bla"
                     /label=AmpR promoter
     CDS             complement(2785..3867)
                     /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     3829..3848
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(3868..3945)
                     /gene="lacI"
                     /label=lacI promoter
     promoter        3991..4009
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    4010..4034
                     /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             4090..4107
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
ORIGIN
        1 ggcagcacgc aactttttga agacaacaaa gcaagtgaac agagcgagca atttacacaa
       61 ctaaaaatta gcgagtggat tgttgactat atgtcagatt tactagaaat tgaacccgat
      121 gaagtagatg tagaaactac tttcgcgcgt tatggtttag attcatcagc cgcagtgatt
      181 ttaacgggag atttaggaaa ttggttagga aaagagatag aaccaacagt catgtatgat
      241 tacccaacaa tctccaagtt agcagaatat gtggctgagg aaaactaata actgcaggtc
      301 gacaagcttg cggccgcata atgcttaagt cgaacagaaa gtaatcgtat tgtacacggc
      361 cgcataatcg aaattaatac gactcactat aggggaattg tgagcggata acaattcccc
      421 atcttagtat attagttaag tataagaagg agatatacat atggcagatc tcaattggat
      481 atcggccggc cacgcgatcg ctgacgtcgg taccctcgag tctggtaaag aaaccgctgc
      541 tgcgaaattt gaacgccagc acatggactc gtctactagc gcagcttaat taacctaggc
      601 tgctgccacc gctgagcaat aactagcata accccttggg gcctctaaac gggtcttgag
      661 gggttttttg ctgaaacctc aggcatttga gaagcacacg gtcacactgc ttccggtagt
      721 caataaaccg gtaaaccagc aatagacata agcggctatt taacgaccct gccctgaacc
      781 gacgaccggg tcatcgtggc cggatcttgc ggcccctcgg cttgaacgaa ttgttagaca
      841 ttatttgccg actaccttgg tgatctcgcc tttcacgtag tggacaaatt cttccaactg
      901 atctgcgcgc gaggccaagc gatcttcttc ttgtccaaga taagcctgtc tagcttcaag
      961 tatgacgggc tgatactggg ccggcaggcg ctccattgcc cagtcggcag cgacatcctt
     1021 cggcgcgatt ttgccggtta ctgcgctgta ccaaatgcgg gacaacgtaa gcactacatt
     1081 tcgctcatcg ccagcccagt cgggcggcga gttccatagc gttaaggttt catttagcgc
     1141 ctcaaataga tcctgttcag gaaccggatc aaagagttcc tccgccgctg gacctaccaa
     1201 ggcaacgcta tgttctcttg cttttgtcag caagatagcc agatcaatgt cgatcgtggc
     1261 tggctcgaag atacctgcaa gaatgtcatt gcgctgccat tctccaaatt gcagttcgcg
     1321 cttagctgga taacgccacg gaatgatgtc gtcgtgcaca acaatggtga cttctacagc
     1381 gcggagaatc tcgctctctc caggggaagc cgaagtttcc aaaaggtcgt tgatcaaagc
     1441 tcgccgcgtt gtttcatcaa gccttacggt caccgtaacc agcaaatcaa tatcactgtg
     1501 tggcttcagg ccgccatcca ctgcggagcc gtacaaatgt acggccagca acgtcggttc
     1561 gagatggcgc tcgatgacgc caactacctc tgatagttga gtcgatactt cggcgatcac
     1621 cgcttccctc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     1681 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagcta gctcactcgg
     1741 tcgctacgct ccgggcgtga gactgcggcg ggcgctgcgg acacatacaa agttacccac
     1801 agattccgtg gataagcagg ggactaacat gtgaggcaaa acagcagggc cgcgccggtg
     1861 gcgtttttcc ataggctccg ccctcctgcc agagttcaca taaacagacg cttttccggt
     1921 gcatctgtgg gagccgtgag gctcaaccat gaatctgaca gtacgggcga aacccgacag
     1981 gacttaaaga tccccaccgt ttccggcggg tcgctccctc ttgcgctctc ctgttccgac
     2041 cctgccgttt accggatacc tgttccgcct ttctccctta cgggaagtgt ggcgctttct
     2101 catagctcac acactggtat ctcggctcgg tgtaggtcgt tcgctccaag ctgggctgta
     2161 agcaagaact ccccgttcag cccgactgct gcgccttatc cggtaactgt tcacttgagt
     2221 ccaacccgga aaagcacggt aaaacgccac tggcagcagc cattggtaac tgggagttcg
     2281 cagaggattt gtttagctaa acacgcggtt gctcttgaag tgtgcgccaa agtccggcta
     2341 cactggaagg acagatttgg ttgctgtgct ctgcgaaagc cagttaccac ggttaagcag
     2401 ttccccaact gacttaacct tcgatcaaac cacctcccca ggtggttttt tcgaaggtta
     2461 agtcagttgg ggaactgctt aaccttcgat caaaccacct ccccaggtgg ttttttcgtt
     2521 tacagggcaa aagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     2581 tactgaaccg ctctagattt cagtgcaatt tatctcttca aatgtagcac ctgaagtcag
     2641 ccccatacga tataagttgt aattctcatg ttagtcatgc cccgcgccca ccggaaggag
     2701 ctgactgggt tgaaggctct caagggcatc ggtcgagatc ccggtgccta atgagtgagc
     2761 taacttacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     2821 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgccag
     2881 ggtggttttt cttttcacca gtgagacggg caacagctga ttgcccttca ccgcctggcc
     2941 ctgagagagt tgcagcaagc ggtccacgct ggtttgcccc agcaggcgaa aatcctgttt
     3001 gatggtggtt aacggcggga tataacatga gctgtcttcg gtatcgtcgt atcccactac
     3061 cgagatgtcc gcaccaacgc gcagcccgga ctcggtaatg gcgcgcattg cgcccagcgc
     3121 catctgatcg ttggcaacca gcatcgcagt gggaacgatg ccctcattca gcatttgcat
     3181 ggtttgttga aaaccggaca tggcactcca gtcgccttcc cgttccgcta tcggctgaat
     3241 ttgattgcga gtgagatatt tatgccagcc agccagacgc agacgcgccg agacagaact
     3301 taatgggccc gctaacagcg cgatttgctg gtgacccaat gcgaccagat gctccacgcc
     3361 cagtcgcgta ccgtcttcat gggagaaaat aatactgttg atgggtgtct ggtcagagac
     3421 atcaagaaat aacgccggaa cattagtgca ggcagcttcc acagcaatgg catcctggtc
     3481 atccagcgga tagttaatga tcagcccact gacgcgttgc gcgagaagat tgtgcaccgc
     3541 cgctttacag gcttcgacgc cgcttcgttc taccatcgac accaccacgc tggcacccag
     3601 ttgatcggcg cgagatttaa tcgccgcgac aatttgcgac ggcgcgtgca gggccagact
     3661 ggaggtggca acgccaatca gcaacgactg tttgcccgcc agttgttgtg ccacgcggtt
     3721 gggaatgtaa ttcagctccg ccatcgccgc ttccactttt tcccgcgttt tcgcagaaac
     3781 gtggctggcc tggttcacca cgcgggaaac ggtctgataa gagacaccgg catactctgc
     3841 gacatcgtat aacgttactg gtttcacatt caccaccctg aattgactct cttccgggcg
     3901 ctatcatgcc ataccgcgaa aggttttgcg ccattcgatg gtgtccggga tctcgacgct
     3961 ctcccttatg cgactcctgc attaggaaat taatacgact cactataggg gaattgtgag
     4021 cggataacaa ttcccctgta gaaataattt tgtttaactt taataaggag atataccatg
     4081 ggcagcagcc atcaccatca tcaccac
//
