LOCUS       sequence-161677-        6721 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses nadV from Haemophilus ducreyi in Escherichia coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6721)
  AUTHORS   Marinescu GC, Popescu RG, Dinischiotu A
  TITLE     Size Exclusion Chromatography Method for Purification of 
            Nicotinamide Mononucleotide (NMN) from Bacterial Cells.
  JOURNAL   Sci Rep. 2018 Mar 13;8(1):4433. doi: 10.1038/s41598-018-22806-8.
  PUBMED    29535407
REFERENCE   2  (bases 1 to 6721)
  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: "2018-03-13"; volume: "8(1):4433. doi"; pages: " 
            10.1038/s41598-018-22806-8"
COMMENT     Sequence Label: pET28a-hdNadV
FEATURES             Location/Qualifiers
     source          1..6721
                     /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             1658..1680
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    1695..1719
                     /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(1719..1738)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1720..1738)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1822..1841)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        2047..2124
                     /gene="lacI"
                     /label=lacI promoter
     CDS             2125..3207
                     /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(2144..2163)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             4016..4207
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(4201..4223)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    4309..4451
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     4364..4383
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4466..4483)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4637..5225)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4717..4736)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(5336..5355)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             5347..6162
                     /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             5347..6162
                     /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(5420..5439)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      complement(6255..6710)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6392..6413)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6604..6623
                     /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 gagtcacagg gtcgtgcggc
      181 taaccagcag gttctgacgg atttcatcaa agtcggtctg acgcagcagt ttaccatctt
      241 cgaacagcag ttccagcagg tcatctgaaa agtcatcggc ggacgtcagg ccgtccacat
      301 aggtatcgcg gctcagaact ttcacacgac ctttctgcga ttttttgaag ccgtcatcgg
      361 ttttcgggtt tttgaagatg gctttttctt caccgttgat cgtaatgctg gtagctttca
      421 gtgcaaagcc cagcgtatcg cgggtattac gttggtaggt ctgcgcaccc acgccgaaga
      481 cgatgttact agaggcaaaa cctttcgctt gcaggccttc caggattttg aacatttttt
      541 cgtacgtaac accgtcgcca tagattgcac caatgtgcgg attgataact ttatagcctt
      601 tctggttcac ggtaccgcca aagatatccc acagacattc aatcagacct ttgcgttcat
      661 gttccgtgtc ggcggtcgga tcaccgcaga taatcgcgaa aaagttgccg ctatccgggc
      721 gaatcaccag acgagcgttt tccggacgtg cgataatttc ctgtttcagc agcggcagat
      781 tcacggtgat gttgtgccaa aagtccgtgg tatcactgac aatagacagc atgttatgcg
      841 ggaatttagc catcagatag cgaaacgtgc tcagttcatc gacgccgtgg ctcgacataa
      901 cgctatgttc cgatgccggg atgctggtac caatcagtga ggaactgccg taataggctt
      961 caacgaatga cagcgccgga atcgtatcgg tacccagaaa ggacgtcaga tgaccggcgc
     1021 cgctggtttc cgccgattcc agagagctca tgccgcgcat tgagaagtcg tgggattgaa
     1081 acggcacatg ttcctgatta tcacacgttt cgttagcgaa tttgatcagt gcggtacggt
     1141 aggcaaacgc aattgacgcg ctggtcatcg gctgccacag actgacattg atcagcgttt
     1201 ccagatagtt ggtcagccaa aagaaatcag aatgcgtatt ttcgatggtc atgaccggaa
     1261 ctttaatagc aacggtttta ccttccggaa ttgctttgat acgaatcggc aggtaaccca
     1321 gttcgtgcag tttggcgata tgttcgcccg tgtcttccag ctgcagggtt ttttcaataa
     1381 acgcactata ttcggtcacg acgtcgtgtt tatcacggct gaaaaagtta tcgttgaagt
     1441 aatggatcag gtatttgatg atgaaagctt ggaagccgaa gctaaccact tgggtcaggt
     1501 acggtgcctg ttcgttcgaa cgcggcgtaa aggtactata gataatctga gagcattccg
     1561 gatacataat acgatgagag gttttgtaga agtcgcacag cagggacggg attgccgagg
     1621 cgcgtgagga gtagttcagc agattgtcca tggtatatct ccttcttaaa gttaaacaaa
     1681 attatttcta gaggggaatt gttatccgct cacaattccc ctatagtgag tcgtattaat
     1741 ttcgcgggat cgagatctcg atcctctacg ccggacgcat cgtggccggc atcaccggcg
     1801 ccacaggtgc ggttgctggc gcctatatcg ccgacatcac cgatggggaa gatcgggctc
     1861 gccacttcgg gctcatgagc gcttgtttcg gcgtgggtat ggtggcaggc cccgtggccg
     1921 ggggactgtt gggcgccatc tccttgcatg caccattcct tgcggcggcg gtgctcaacg
     1981 gcctcaacct actactgggc tgcttcctaa tgcaggagtc gcataaggga gagcgtcgag
     2041 atcccggaca ccatcgaatg gcgcaaaacc tttcgcggta tggcatgata gcgcccggaa
     2101 gagagtcaat tcagggtggt gaatgtgaaa ccagtaacgt tatacgatgt cgcagagtat
     2161 gccggtgtct cttatcagac cgtttcccgc gtggtgaacc aggccagcca cgtttctgcg
     2221 aaaacgcggg aaaaagtgga agcggcgatg gcggagctga attacattcc caaccgcgtg
     2281 gcacaacaac tggcgggcaa acagtcgttg ctgattggcg ttgccacctc cagtctggcc
     2341 ctgcacgcgc cgtcgcaaat tgtcgcggcg attaaatctc gcgccgatca actgggtgcc
     2401 agcgtggtgg tgtcgatggt agaacgaagc ggcgtcgaag cctgtaaagc ggcggtgcac
     2461 aatcttctcg cgcaacgcgt cagtgggctg atcattaact atccgctgga tgaccaggat
     2521 gccattgctg tggaagctgc ctgcactaat gttccggcgt tatttcttga tgtctctgac
     2581 cagacaccca tcaacagtat tattttctcc catgaagacg gtacgcgact gggcgtggag
     2641 catctggtcg cattgggtca ccagcaaatc gcgctgttag cgggcccatt aagttctgtc
     2701 tcggcgcgtc tgcgtctggc tggctggcat aaatatctca ctcgcaatca aattcagccg
     2761 atagcggaac gggaaggcga ctggagtgcc atgtccggtt ttcaacaaac catgcaaatg
     2821 ctgaatgagg gcatcgttcc cactgcgatg ctggttgcca acgatcagat ggcgctgggc
     2881 gcaatgcgcg ccattaccga gtccgggctg cgcgttggtg cggatatctc ggtagtggga
     2941 tacgacgata ccgaagacag ctcatgttat atcccgccgt taaccaccat caaacaggat
     3001 tttcgcctgc tggggcaaac cagcgtggac cgcttgctgc aactctctca gggccaggcg
     3061 gtgaagggca atcagctgtt gcccgtctca ctggtgaaaa gaaaaaccac cctggcgccc
     3121 aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag
     3181 gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtaagtt agctcactca
     3241 ttaggcaccg ggatctcgac cgatgccctt gagagccttc aacccagtca gctccttccg
     3301 gtgggcgcgg ggcatgacta tcgtcgccgc acttatgact gtcttcttta tcatgcaact
     3361 cgtaggacag gtgccggcag cgctctgggt cattttcggc gaggaccgct ttcgctggag
     3421 cgcgacgatg atcggcctgt cgcttgcggt attcggaatc ttgcacgccc tcgctcaagc
     3481 cttcgtcact ggtcccgcca ccaaacgttt cggcgagaag caggccatta tcgccggcat
     3541 ggcggcccca cgggtgcgca tgatcgtgct cctgtcgttg aggacccggc taggctggcg
     3601 gggttgcctt actggttagc agaatgaatc accgatacgc gagcgaacgt gaagcgactg
     3661 ctgctgcaaa acgtctgcga cctgagcaac aacatgaatg gtcttcggtt tccgtgtttc
     3721 gtaaagtctg gaaacgcgga agtcagcgcc ctgcaccatt atgttccgga tctgcatcgc
     3781 aggatgctgc tggctaccct gtggaacacc tacatctgta ttaacgaagc gctggcattg
     3841 accctgagtg atttttctct ggtcccgccg catccatacc gccagttgtt taccctcaca
     3901 acgttccagt aaccgggcat gttcatcatc agtaacccgt atcgtgagca tcctctctcg
     3961 tttcatcggt atcattaccc ccatgaacag aaatccccct tacacggagg catcagtgac
     4021 caaacaggaa aaaaccgccc ttaacatggc ccgctttatc agaagccaga cattaacgct
     4081 tctggagaaa ctcaacgagc tggacgcgga tgaacaggca gacatctgtg aatcgcttca
     4141 cgaccacgct gatgagcttt accgcagctg cctcgcgcgt ttcggtgatg acggtgaaaa
     4201 cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag
     4261 cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggcg cagccatgac
     4321 ccagtcacgt agcgatagcg gagtgtatac tggcttaact atgcggcatc agagcagatt
     4381 gtactgagag tgcaccatat atgcggtgtg aaataccgca cagatgcgta aggagaaaat
     4441 accgcatcag gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     4501 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     4561 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     4621 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     4681 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     4741 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     4801 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     4861 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     4921 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     4981 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     5041 tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc
     5101 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     5161 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     5221 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     5281 gttaagggat tttggtcatg aacaataaaa ctgtctgctt acataaacag taatacaagg
     5341 ggtgttatga gccatattca acgggaaacg tcttgctcta ggccgcgatt aaattccaac
     5401 atggatgctg atttatatgg gtataaatgg gctcgcgata atgtcgggca atcaggtgcg
     5461 acaatctatc gattgtatgg gaagcccgat gcgccagagt tgtttctgaa acatggcaaa
     5521 ggtagcgttg ccaatgatgt tacagatgag atggtcagac taaactggct gacggaattt
     5581 atgcctcttc cgaccatcaa gcattttatc cgtactcctg atgatgcatg gttactcacc
     5641 actgcgatcc ccgggaaaac agcattccag gtattagaag aatatcctga ttcaggtgaa
     5701 aatattgttg atgcgctggc agtgttcctg cgccggttgc attcgattcc tgtttgtaat
     5761 tgtcctttta acagcgatcg cgtatttcgt ctcgctcagg cgcaatcacg aatgaataac
     5821 ggtttggttg atgcgagtga ttttgatgac gagcgtaatg gctggcctgt tgaacaagtc
     5881 tggaaagaaa tgcataaact tttgccattc tcaccggatt cagtcgtcac tcatggtgat
     5941 ttctcacttg ataaccttat ttttgacgag gggaaattaa taggttgtat tgatgttgga
     6001 cgagtcggaa tcgcagaccg ataccaggat cttgccatcc tatggaactg cctcggtgag
     6061 ttttctcctt cattacagaa acggcttttt caaaaatatg gtattgataa tcctgatatg
     6121 aataaattgc agtttcattt gatgctcgat gagtttttct aagaattaat tcatgagcgg
     6181 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     6241 aaaagtgcca cctgaaattg taaacgttaa tattttgtta aaattcgcgt taaatttttg
     6301 ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa
     6361 agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa
     6421 gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg
     6481 tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa
     6541 ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa
     6601 ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct
     6661 gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt cccattcgcc
     6721 a
//
