LOCUS       sequence-194125-        7702 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pET28a-baPrs-hdNadV is a bicistronic vector designed for the 
            simultaneous expression of PRS from Bacillus amyloliquefaciens and 
            NadV from Haemophilus ducreyi.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7702)
  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 7702)
  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-baPrs-hdNadV
FEATURES             Location/Qualifiers
     source          1..7702
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(31..78)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     74..92
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(145..162)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             2648..2670
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    2685..2709
                     /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(2709..2728)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2710..2728)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2812..2831)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        3037..3114
                     /gene="lacI"
                     /label=lacI promoter
     CDS             3115..4197
                     /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(3134..3153)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             5006..5197
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(5191..5213)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    5299..5438
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     5354..5373
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5453..5470)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5624..6212)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5704..5723)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(6323..6342)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             6334..7149
                     /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             6334..7149
                     /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(6407..6426)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      complement(7241..7696)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7378..7399)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7590..7609
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 cgccaatccg gatatagttc ctcctttcag caaaaaaccc ctcaagaccc gtttagaggc
       61 cccaaggggt tatgctagtt attgctcagc ggtggcagca gccaactcag cttcctttcg
      121 ggctttgtta gcagccggat ctcagtggtg gtggtggtgg tgctcgagtc acagggtcgt
      181 gcggctaacc agcaggttct gacggatttc atcaaagtcg gtctgacgca gcagtttacc
      241 atcttcgaac agcagttcca gcaggtcatc tgaaaagtca tcggcggacg tcaggccgtc
      301 cacataggta tcgcggctca gaactttcac acgacctttc tgcgattttt tgaagccgtc
      361 atcggttttc gggtttttga agatggcttt ttcttcaccg ttgatcgtaa tgctggtagc
      421 tttcagtgca aagcccagcg tatcgcgggt attacgttgg taggtctgcg cacccacgcc
      481 gaagacgatg ttactagagg caaaaccttt cgcttgcagg ccttccagga ttttgaacat
      541 tttttcgtac gtaacaccgt cgccatagat tgcaccaatg tgcggattga taactttata
      601 gcctttctgg ttcacggtac cgccaaagat atcccacaga cattcaatca gacctttgcg
      661 ttcatgttcc gtgtcggcgg tcggatcacc gcagataatc gcgaaaaagt tgccgctatc
      721 cgggcgaatc accagacgag cgttttccgg acgtgcgata atttcctgtt tcagcagcgg
      781 cagattcacg gtgatgttgt gccaaaagtc cgtggtatca ctgacaatag acagcatgtt
      841 atgcgggaat ttagccatca gatagcgaaa cgtgctcagt tcatcgacgc cgtggctcga
      901 cataacgcta tgttccgatg ccgggatgct ggtaccaatc agtgaggaac tgccgtaata
      961 ggcttcaacg aatgacagcg ccggaatcgt atcggtaccc agaaaggacg tcagatgacc
     1021 ggcgccgctg gtttccgccg attccagaga gctcatgccg cgcattgaga agtcgtggga
     1081 ttgaaacggc acatgttcct gattatcaca cgtttcgtta gcgaatttga tcagtgcggt
     1141 acggtaggca aacgcaattg acgcgctggt catcggctgc cacagactga cattgatcag
     1201 cgtttccaga tagttggtca gccaaaagaa atcagaatgc gtattttcga tggtcatgac
     1261 cggaacttta atagcaacgg ttttaccttc cggaattgct ttgatacgaa tcggcaggta
     1321 acccagttcg tgcagtttgg cgatatgttc gcccgtgtct tccagctgca gggttttttc
     1381 aataaacgca ctatattcgg tcacgacgtc gtgtttatca cggctgaaaa agttatcgtt
     1441 gaagtaatgg atcaggtatt tgatgatgaa agcttggaag ccgaagctaa ccacttgggt
     1501 caggtacggt gcctgttcgt tcgaacgcgg cgtaaaggta ctatagataa tctgagagca
     1561 ttccggatac ataatacgat gagaggtttt gtagaagtcg cacagcaggg acgggattgc
     1621 cgaggcgcgt gaggagtagt tcagcagatt gtccatggtt actattcggt ttaaactttc
     1681 ggtttagctg aataaatagc tgacagattg tttctcgtga acgcggataa tcgcttcagc
     1741 cagaagcggt ccgacagaaa gctgtttaaa gcgttcgatt tttttctcct caggaagctt
     1801 aatgctgttc gtcacgacaa gctccttgat tttagagtta ttgatccgct caacggcagg
     1861 gcctgacaat acagggtgcg tacagcatgc atacacttcc gcagctccgt tttcaaccag
     1921 cgcgtttgcg gcaagtgtga ttgtgccggc cgtatcaata atatcatcga tcagaatcgc
     1981 tgtttttcct tccacgttac cgacgatgtt catcacttct acctcattcg gtctaggacg
     2041 gcgtttgtcg ataatcgcga tcggcgcttt cagacggtcc gcaagtttgc gggcgcgcgt
     2101 cacaccgccg tggtcaggtg aaacaataac aatatctttc aagtctttac cttcaaaata
     2161 atggcctaaa atcggaacac ccattaagtg gtcaatcgga atatcaaaga aaccttgaat
     2221 ttgcggcgcg tgtatatcaa gtgcgattac gcgtgtcgct cccgctgttt caagcaggtt
     2281 tgcaaaaagt tttgccgtaa tcggttcacg tgatcttgct tttctgtcct ggcgcgcata
     2341 accgtagtac gggatgacaa tgttgattgt tttcgcggaa gcgcgcttta aagcgtcaac
     2401 cataatcagc aattccatga tgtgctcgtt gaccggcgca cttgtagact ggatgatgta
     2461 gcaatcacag ccgcgaatac tttcttctat gttgatctgg acctctccgt cactaaatct
     2521 tgtaacagag cactttccta attgaactcc tacattatcc gcgatttctt ttgcaagttc
     2581 tggatttgaa ttcaaagaaa aaatctttaa attcttatct ccgtattcgt tagacatcgt
     2641 gcctatatct ccttcttaaa gttaaacaaa attatttcta gaggggaatt gttatccgct
     2701 cacaattccc ctatagtgag tcgtattaat ttcgcgggat cgagatctcg atcctctacg
     2761 ccggacgcat cgtggccggc atcaccggcg ccacaggtgc ggttgctggc gcctatatcg
     2821 ccgacatcac cgatggggaa gatcgggctc gccacttcgg gctcatgagc gcttgtttcg
     2881 gcgtgggtat ggtggcaggc cccgtggccg ggggactgtt gggcgccatc tccttgcatg
     2941 caccattcct tgcggcggcg gtgctcaacg gcctcaacct actactgggc tgcttcctaa
     3001 tgcaggagtc gcataaggga gagcgtcgag atcccggaca ccatcgaatg gcgcaaaacc
     3061 tttcgcggta tggcatgata gcgcccggaa gagagtcaat tcagggtggt gaatgtgaaa
     3121 ccagtaacgt tatacgatgt cgcagagtat gccggtgtct cttatcagac cgtttcccgc
     3181 gtggtgaacc aggccagcca cgtttctgcg aaaacgcggg aaaaagtgga agcggcgatg
     3241 gcggagctga attacattcc caaccgcgtg gcacaacaac tggcgggcaa acagtcgttg
     3301 ctgattggcg ttgccacctc cagtctggcc ctgcacgcgc cgtcgcaaat tgtcgcggcg
     3361 attaaatctc gcgccgatca actgggtgcc agcgtggtgg tgtcgatggt agaacgaagc
     3421 ggcgtcgaag cctgtaaagc ggcggtgcac aatcttctcg cgcaacgcgt cagtgggctg
     3481 atcattaact atccgctgga tgaccaggat gccattgctg tggaagctgc ctgcactaat
     3541 gttccggcgt tatttcttga tgtctctgac cagacaccca tcaacagtat tattttctcc
     3601 catgaagacg gtacgcgact gggcgtggag catctggtcg cattgggtca ccagcaaatc
     3661 gcgctgttag cgggcccatt aagttctgtc tcggcgcgtc tgcgtctggc tggctggcat
     3721 aaatatctca ctcgcaatca aattcagccg atagcggaac gggaaggcga ctggagtgcc
     3781 atgtccggtt ttcaacaaac catgcaaatg ctgaatgagg gcatcgttcc cactgcgatg
     3841 ctggttgcca acgatcagat ggcgctgggc gcaatgcgcg ccattaccga gtccgggctg
     3901 cgcgttggtg cggatatctc ggtagtggga tacgacgata ccgaagacag ctcatgttat
     3961 atcccgccgt taaccaccat caaacaggat tttcgcctgc tggggcaaac cagcgtggac
     4021 cgcttgctgc aactctctca gggccaggcg gtgaagggca atcagctgtt gcccgtctca
     4081 ctggtgaaaa gaaaaaccac cctggcgccc aatacgcaaa ccgcctctcc ccgcgcgttg
     4141 gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg
     4201 caacgcaatt aatgtaagtt agctcactca ttaggcaccg ggatctcgac cgatgccctt
     4261 gagagccttc aacccagtca gctccttccg gtgggcgcgg ggcatgacta tcgtcgccgc
     4321 acttatgact gtcttcttta tcatgcaact cgtaggacag gtgccggcag cgctctgggt
     4381 cattttcggc gaggaccgct ttcgctggag cgcgacgatg atcggcctgt cgcttgcggt
     4441 attcggaatc ttgcacgccc tcgctcaagc cttcgtcact ggtcccgcca ccaaacgttt
     4501 cggcgagaag caggccatta tcgccggcat ggcggcccca cgggtgcgca tgatcgtgct
     4561 cctgtcgttg aggacccggc taggctggcg gggttgcctt actggttagc agaatgaatc
     4621 accgatacgc gagcgaacgt gaagcgactg ctgctgcaaa acgtctgcga cctgagcaac
     4681 aacatgaatg gtcttcggtt tccgtgtttc gtaaagtctg gaaacgcgga agtcagcgcc
     4741 ctgcaccatt atgttccgga tctgcatcgc aggatgctgc tggctaccct gtggaacacc
     4801 tacatctgta ttaacgaagc gctggcattg accctgagtg atttttctct ggtcccgccg
     4861 catccatacc gccagttgtt taccctcaca acgttccagt aaccgggcat gttcatcatc
     4921 agtaacccgt atcgtgagca tcctctctcg tttcatcggt atcattaccc ccatgaacag
     4981 aaatccccct tacacggagg catcagtgac caaacaggaa aaaaccgccc ttaacatggc
     5041 ccgctttatc agaagccaga cattaacgct tctggagaaa ctcaacgagc tggacgcgga
     5101 tgaacaggca gacatctgtg aatcgcttca cgaccacgct gatgagcttt accgcagctg
     5161 cctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt
     5221 cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg
     5281 tgttggcggg tgtcggggcg cagccatgac ccagtcacgt agcgatagcg gagtgtatac
     5341 tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccattg cggtgtgaaa
     5401 taccgcacag atgcgtaagg agaaaatacc gcatcaggcg ctcttccgct tcctcgctca
     5461 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     5521 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     5581 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     5641 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     5701 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     5761 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     5821 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     5881 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     5941 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     6001 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     6061 gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     6121 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     6181 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     6241 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaac aataaaactg
     6301 tctgcttaca taaacagtaa tacaaggggt gttatgagcc atattcaacg ggaaacgtct
     6361 tgctctaggc cgcgattaaa ttccaacatg gatgctgatt tatatgggta taaatgggct
     6421 cgcgataatg tcgggcaatc aggtgcgaca atctatcgat tgtatgggaa gcccgatgcg
     6481 ccagagttgt ttctgaaaca tggcaaaggt agcgttgcca atgatgttac agatgagatg
     6541 gtcagactaa actggctgac ggaatttatg cctcttccga ccatcaagca ttttatccgt
     6601 actcctgatg atgcatggtt actcaccact gcgatccccg ggaaaacagc attccaggta
     6661 ttagaagaat atcctgattc aggtgaaaat attgttgatg cgctggcagt gttcctgcgc
     6721 cggttgcatt cgattcctgt ttgtaattgt ccttttaaca gcgatcgcgt atttcgtctc
     6781 gctcaggcgc aatcacgaat gaataacggt ttggttgatg cgagtgattt tgatgacgag
     6841 cgtaatggct ggcctgttga acaagtctgg aaagaaatgc ataaactttt gccattctca
     6901 ccggattcag tcgtcactca tggtgatttc tcacttgata accttatttt tgacgagggg
     6961 aaattaatag gttgtattga tgttggacga gtcggaatcg cagaccgata ccaggatctt
     7021 gccatcctat ggaactgcct cggtgagttt tctccttcat tacagaaacg gctttttcaa
     7081 aaatatggta ttgataatcc tgatatgaat aaattgcagt ttcatttgat gctcgatgag
     7141 tttttctaag aattaattca tgagcggata catatttgaa tgtatttaga aaaataaaca
     7201 aataggggtt ccgcgcacat ttccccgaaa agtgccacct aaattgtaag cgttaatatt
     7261 ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa
     7321 atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca
     7381 gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc
     7441 gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg
     7501 aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg
     7561 ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg
     7621 gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg
     7681 ccgctacagg gcgcgtccca tt
//
