LOCUS       sequence-161675-        6709 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses nadV from Shewanella oneidensis in Escherichia coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6709)
  AUTHORS   Marinescu GC, Popescu RG, Stoian G, Dinischiotu A
  TITLE     beta-nicotinamide mononucleotide (NMN) production in Escherichia 
            coli.
  JOURNAL   Sci Rep. 2018 Aug 16;8(1):12278. doi: 10.1038/s41598-018-30792-0.
  PUBMED    30115969
REFERENCE   2  (bases 1 to 6709)
  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: "Sci Rep."; 
            date: "2018-08-16"; volume: "8(1):12278. doi"; pages: " 
            10.1038/s41598-018-30792-0"
COMMENT     Sequence Label: pET28a-soNadV
FEATURES             Location/Qualifiers
     source          1..6709
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(26..73)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     69..87
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(140..157)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             1646..1668
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    1683..1707
                     /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)."
     primer_bind     complement(1707..1726)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1708..1726)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1810..1829)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        2035..2112
                     /gene="lacI"
                     /label=lacI promoter
     CDS             2113..3195
                     /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     complement(2132..2151)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             4004..4195
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(4189..4211)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    4297..4439
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     4352..4371
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4454..4471)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4625..5213)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4705..4724)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(5324..5343)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             5335..6150
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             5335..6150
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(5408..5427)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      complement(6243..6698)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6380..6401)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6592..6611
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 atccggatat agttcctcct ttcagcaaaa aacccctcaa gacccgttta gaggccccaa
       61 ggggttatgc tagttattgc tcagcggtgg cagcagccaa ctcagcttcc tttcgggctt
      121 tgttagcagc cggatctcag tggtggtggt ggtggtgctc gagttagcga cgggatgcgg
      181 ccaggcgggc acggatgtca gccagggagt attcggtcag cagttcacca tcagcgaaga
      241 ccggaacaaa ttcgcaattg ttcacgtctt gccacgatgc ctgatctttc agatagaatt
      301 ggccctgttc atctttaaac acggcgacca gacctttcgc tgattttttg gtgccgtcat
      361 ccgttttcgg atctttgaaa atttcgcggt ctttaccatc aactttcgca aaggtagctt
      421 tcactgcgta gccgtcggta tcacgcgtca cgtgctgata ggtgaacgaa ccgatgccaa
      481 agacaatgtt cgttgaagca aaaccttttg ctttcaggcc ggcacagata gcttctgcgc
      541 gttcgatggt aatggagtca ccgtaaatca ggccaacatg actatccagc agtttataac
      601 ctttcgccgt ggtagaaccg ccaaagacat cccacaggca ttcgatggca cctttgtatt
      661 ccgggctacc aatcggggcc tgcgggtcac cacagataat tttaaccggg cagccggtat
      721 ccggacgaaa aaccacttta ccgtcgcgag ccaggatacg atctttcagt gccagggtga
      781 attccgtcat cacctgccaa aagtcccaag aatcgctaac gatactcaca atgccggtcg
      841 gatagatatc ttcaatcagg cgacgaaacg tttccagttc attttccata ccgccggcac
      901 acataaccga gtgttcggtc gccggcactg aaccgccgac cagttctttg tcagaatcag
      961 ccaggtaata ctgttccagg aagtcgattg ccggaatggt atccgtaccg gtaaaggaca
     1021 gcaggtgagc tgcgccactc ataatggccg cttcgacacc gtacatgccg cggaagctaa
     1081 agtcatgacc ttgccaatca acgaatgcga tgtcgccatc ggtttccagg gcgtacgctt
     1141 caaaaatttt acgatactgc agggccgtgg tcgcagacgt acacatgccc cacacattgg
     1201 cgctcaggat ggtttccaga tagttcgtca gccaaaagaa acgttcatcg gtgttgtaca
     1261 gaatcaggca cgggactttc agcggataaa cgctaccttc cggcagggcc atgattttaa
     1321 tcggcaggta gcccagatcg tgcagttctt caatatgctg cgtaccaaca ttgttcgggc
     1381 ccagataatt gttgatgcga cgggagaatt ggcaaatcac ttgttctttc ggttgctgga
     1441 aaaagtcgcg attccaacta tcgatcaggt aatgtttgat gaagtattgc agaccgaaaa
     1501 acaccatctg atcggtatag ccgcgacggc ttttacgcgc cgtcaggttt gagaagataa
     1561 cttgggtatt gtccgggtat tggcggcgat ggtcaacttt gtagccgtcg attgccgtca
     1621 ccgggttcag ataagccatg gtatatctcc ttcttaaagt taaacaaaat tatttctaga
     1681 ggggaattgt tatccgctca caattcccct atagtgagtc gtattaattt cgcgggatcg
     1741 agatctcgat cctctacgcc ggacgcatcg tggccggcat caccggcgcc acaggtgcgg
     1801 ttgctggcgc ctatatcgcc gacatcaccg atggggaaga tcgggctcgc cacttcgggc
     1861 tcatgagcgc ttgtttcggc gtgggtatgg tggcaggccc cgtggccggg ggactgttgg
     1921 gcgccatctc cttgcatgca ccattccttg cggcggcggt gctcaacggc ctcaacctac
     1981 tactgggctg cttcctaatg caggagtcgc ataagggaga gcgtcgagat cccggacacc
     2041 atcgaatggc gcaaaacctt tcgcggtatg gcatgatagc gcccggaaga gagtcaattc
     2101 agggtggtga atgtgaaacc agtaacgtta tacgatgtcg cagagtatgc cggtgtctct
     2161 tatcagaccg tttcccgcgt ggtgaaccag gccagccacg tttctgcgaa aacgcgggaa
     2221 aaagtggaag cggcgatggc ggagctgaat tacattccca accgcgtggc acaacaactg
     2281 gcgggcaaac agtcgttgct gattggcgtt gccacctcca gtctggccct gcacgcgccg
     2341 tcgcaaattg tcgcggcgat taaatctcgc gccgatcaac tgggtgccag cgtggtggtg
     2401 tcgatggtag aacgaagcgg cgtcgaagcc tgtaaagcgg cggtgcacaa tcttctcgcg
     2461 caacgcgtca gtgggctgat cattaactat ccgctggatg accaggatgc cattgctgtg
     2521 gaagctgcct gcactaatgt tccggcgtta tttcttgatg tctctgacca gacacccatc
     2581 aacagtatta ttttctccca tgaagacggt acgcgactgg gcgtggagca tctggtcgca
     2641 ttgggtcacc agcaaatcgc gctgttagcg ggcccattaa gttctgtctc ggcgcgtctg
     2701 cgtctggctg gctggcataa atatctcact cgcaatcaaa ttcagccgat agcggaacgg
     2761 gaaggcgact ggagtgccat gtccggtttt caacaaacca tgcaaatgct gaatgagggc
     2821 atcgttccca ctgcgatgct ggttgccaac gatcagatgg cgctgggcgc aatgcgcgcc
     2881 attaccgagt ccgggctgcg cgttggtgcg gatatctcgg tagtgggata cgacgatacc
     2941 gaagacagct catgttatat cccgccgtta accaccatca aacaggattt tcgcctgctg
     3001 gggcaaacca gcgtggaccg cttgctgcaa ctctctcagg gccaggcggt gaagggcaat
     3061 cagctgttgc ccgtctcact ggtgaaaaga aaaaccaccc tggcgcccaa tacgcaaacc
     3121 gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg
     3181 gaaagcgggc agtgagcgca acgcaattaa tgtaagttag ctcactcatt aggcaccggg
     3241 atctcgaccg atgcccttga gagccttcaa cccagtcagc tccttccggt gggcgcgggg
     3301 catgactatc gtcgccgcac ttatgactgt cttctttatc atgcaactcg taggacaggt
     3361 gccggcagcg ctctgggtca ttttcggcga ggaccgcttt cgctggagcg cgacgatgat
     3421 cggcctgtcg cttgcggtat tcggaatctt gcacgccctc gctcaagcct tcgtcactgg
     3481 tcccgccacc aaacgtttcg gcgagaagca ggccattatc gccggcatgg cggccccacg
     3541 ggtgcgcatg atcgtgctcc tgtcgttgag gacccggcta ggctggcggg gttgccttac
     3601 tggttagcag aatgaatcac cgatacgcga gcgaacgtga agcgactgct gctgcaaaac
     3661 gtctgcgacc tgagcaacaa catgaatggt cttcggtttc cgtgtttcgt aaagtctgga
     3721 aacgcggaag tcagcgccct gcaccattat gttccggatc tgcatcgcag gatgctgctg
     3781 gctaccctgt ggaacaccta catctgtatt aacgaagcgc tggcattgac cctgagtgat
     3841 ttttctctgg tcccgccgca tccataccgc cagttgttta ccctcacaac gttccagtaa
     3901 ccgggcatgt tcatcatcag taacccgtat cgtgagcatc ctctctcgtt tcatcggtat
     3961 cattaccccc atgaacagaa atccccctta cacggaggca tcagtgacca aacaggaaaa
     4021 aaccgccctt aacatggccc gctttatcag aagccagaca ttaacgcttc tggagaaact
     4081 caacgagctg gacgcggatg aacaggcaga catctgtgaa tcgcttcacg accacgctga
     4141 tgagctttac cgcagctgcc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat
     4201 gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg
     4261 tcagggcgcg tcagcgggtg ttggcgggtg tcggggcgca gccatgaccc agtcacgtag
     4321 cgatagcgga gtgtatactg gcttaactat gcggcatcag agcagattgt actgagagtg
     4381 caccatatat gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc
     4441 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     4501 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     4561 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     4621 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     4681 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     4741 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     4801 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     4861 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     4921 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     4981 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     5041 ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag
     5101 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     5161 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     5221 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     5281 tggtcatgaa caataaaact gtctgcttac ataaacagta atacaagggg tgttatgagc
     5341 catattcaac gggaaacgtc ttgctctagg ccgcgattaa attccaacat ggatgctgat
     5401 ttatatgggt ataaatgggc tcgcgataat gtcgggcaat caggtgcgac aatctatcga
     5461 ttgtatggga agcccgatgc gccagagttg tttctgaaac atggcaaagg tagcgttgcc
     5521 aatgatgtta cagatgagat ggtcagacta aactggctga cggaatttat gcctcttccg
     5581 accatcaagc attttatccg tactcctgat gatgcatggt tactcaccac tgcgatcccc
     5641 gggaaaacag cattccaggt attagaagaa tatcctgatt caggtgaaaa tattgttgat
     5701 gcgctggcag tgttcctgcg ccggttgcat tcgattcctg tttgtaattg tccttttaac
     5761 agcgatcgcg tatttcgtct cgctcaggcg caatcacgaa tgaataacgg tttggttgat
     5821 gcgagtgatt ttgatgacga gcgtaatggc tggcctgttg aacaagtctg gaaagaaatg
     5881 cataaacttt tgccattctc accggattca gtcgtcactc atggtgattt ctcacttgat
     5941 aaccttattt ttgacgaggg gaaattaata ggttgtattg atgttggacg agtcggaatc
     6001 gcagaccgat accaggatct tgccatccta tggaactgcc tcggtgagtt ttctccttca
     6061 ttacagaaac ggctttttca aaaatatggt attgataatc ctgatatgaa taaattgcag
     6121 tttcatttga tgctcgatga gtttttctaa gaattaattc atgagcggat acatatttga
     6181 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     6241 tgaaattgta aacgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
     6301 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
     6361 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
     6421 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
     6481 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
     6541 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
     6601 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
     6661 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgcca
//
