LOCUS       sequence-176682-        7706 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 7706)
  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 7706)
  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..7706
                     /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             2643..2665
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    2680..2704
                     /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(2704..2723)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2705..2723)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2807..2826)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        3032..3109
                     /gene="lacI"
                     /label=lacI promoter
     CDS             3110..4192
                     /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(3129..3148)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             5001..5192
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(5186..5208)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    5294..5436
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     5349..5368
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5451..5468)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5622..6210)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5702..5721)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(6321..6340)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             6332..7147
                     /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             6332..7147
                     /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(6405..6424)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      complement(7240..7695)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7377..7398)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7589..7608
                     /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 tggttactat tcggtttaaa ctttcggttt
     1681 agctgaataa atagctgaca gattgtttct cgtgaacgcg gataatcgct tcagccagaa
     1741 gcggtccgac agaaagctgt ttaaagcgtt cgattttttt ctcctcagga agcttaatgc
     1801 tgttcgtcac gacaagctcc ttgattttag agttattgat ccgctcaacg gcagggcctg
     1861 acaatacagg gtgcgtacag catgcataca cttccgcagc tccgttttca accagcgcgt
     1921 ttgcggcaag tgtgattgtg ccggccgtat caataatatc atcgatcaga atcgctgttt
     1981 ttccttccac gttaccgacg atgttcatca cttctacctc attcggtcta ggacggcgtt
     2041 tgtcgataat cgcgatcggc gctttcagac ggtccgcaag tttgcgggcg cgcgtcacac
     2101 cgccgtggtc aggtgaaaca ataacaatat ctttcaagtc tttaccttca aaataatggc
     2161 ctaaaatcgg aacacccatt aagtggtcaa tcggaatatc aaagaaacct tgaatttgcg
     2221 gcgcgtgtat atcaagtgcg attacgcgtg tcgctcccgc tgtttcaagc aggtttgcaa
     2281 aaagttttgc cgtaatcggt tcacgtgatc ttgcttttct gtcctggcgc gcataaccgt
     2341 agtacgggat gacaatgttg attgttttcg cggaagcgcg ctttaaagcg tcaaccataa
     2401 tcagcaattc catgatgtgc tcgttgaccg gcgcacttgt agactggatg atgtagcaat
     2461 cacagccgcg aatactttct tctatgttga tctggacctc tccgtcacta aatcttgtaa
     2521 cagagcactt tcctaattga actcctacat tatccgcgat ttcttttgca agttctggat
     2581 ttgaattcaa agaaaaaatc tttaaattct tatctccgta ttcgttagac atcgtgccta
     2641 tatctccttc ttaaagttaa acaaaattat ttctagaggg gaattgttat ccgctcacaa
     2701 ttcccctata gtgagtcgta ttaatttcgc gggatcgaga tctcgatcct ctacgccgga
     2761 cgcatcgtgg ccggcatcac cggcgccaca ggtgcggttg ctggcgccta tatcgccgac
     2821 atcaccgatg gggaagatcg ggctcgccac ttcgggctca tgagcgcttg tttcggcgtg
     2881 ggtatggtgg caggccccgt ggccggggga ctgttgggcg ccatctcctt gcatgcacca
     2941 ttccttgcgg cggcggtgct caacggcctc aacctactac tgggctgctt cctaatgcag
     3001 gagtcgcata agggagagcg tcgagatccc ggacaccatc gaatggcgca aaacctttcg
     3061 cggtatggca tgatagcgcc cggaagagag tcaattcagg gtggtgaatg tgaaaccagt
     3121 aacgttatac gatgtcgcag agtatgccgg tgtctcttat cagaccgttt cccgcgtggt
     3181 gaaccaggcc agccacgttt ctgcgaaaac gcgggaaaaa gtggaagcgg cgatggcgga
     3241 gctgaattac attcccaacc gcgtggcaca acaactggcg ggcaaacagt cgttgctgat
     3301 tggcgttgcc acctccagtc tggccctgca cgcgccgtcg caaattgtcg cggcgattaa
     3361 atctcgcgcc gatcaactgg gtgccagcgt ggtggtgtcg atggtagaac gaagcggcgt
     3421 cgaagcctgt aaagcggcgg tgcacaatct tctcgcgcaa cgcgtcagtg ggctgatcat
     3481 taactatccg ctggatgacc aggatgccat tgctgtggaa gctgcctgca ctaatgttcc
     3541 ggcgttattt cttgatgtct ctgaccagac acccatcaac agtattattt tctcccatga
     3601 agacggtacg cgactgggcg tggagcatct ggtcgcattg ggtcaccagc aaatcgcgct
     3661 gttagcgggc ccattaagtt ctgtctcggc gcgtctgcgt ctggctggct ggcataaata
     3721 tctcactcgc aatcaaattc agccgatagc ggaacgggaa ggcgactgga gtgccatgtc
     3781 cggttttcaa caaaccatgc aaatgctgaa tgagggcatc gttcccactg cgatgctggt
     3841 tgccaacgat cagatggcgc tgggcgcaat gcgcgccatt accgagtccg ggctgcgcgt
     3901 tggtgcggat atctcggtag tgggatacga cgataccgaa gacagctcat gttatatccc
     3961 gccgttaacc accatcaaac aggattttcg cctgctgggg caaaccagcg tggaccgctt
     4021 gctgcaactc tctcagggcc aggcggtgaa gggcaatcag ctgttgcccg tctcactggt
     4081 gaaaagaaaa accaccctgg cgcccaatac gcaaaccgcc tctccccgcg cgttggccga
     4141 ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg
     4201 caattaatgt aagttagctc actcattagg caccgggatc tcgaccgatg cccttgagag
     4261 ccttcaaccc agtcagctcc ttccggtggg cgcggggcat gactatcgtc gccgcactta
     4321 tgactgtctt ctttatcatg caactcgtag gacaggtgcc ggcagcgctc tgggtcattt
     4381 tcggcgagga ccgctttcgc tggagcgcga cgatgatcgg cctgtcgctt gcggtattcg
     4441 gaatcttgca cgccctcgct caagccttcg tcactggtcc cgccaccaaa cgtttcggcg
     4501 agaagcaggc cattatcgcc ggcatggcgg ccccacgggt gcgcatgatc gtgctcctgt
     4561 cgttgaggac ccggctaggc tggcggggtt gccttactgg ttagcagaat gaatcaccga
     4621 tacgcgagcg aacgtgaagc gactgctgct gcaaaacgtc tgcgacctga gcaacaacat
     4681 gaatggtctt cggtttccgt gtttcgtaaa gtctggaaac gcggaagtca gcgccctgca
     4741 ccattatgtt ccggatctgc atcgcaggat gctgctggct accctgtgga acacctacat
     4801 ctgtattaac gaagcgctgg cattgaccct gagtgatttt tctctggtcc cgccgcatcc
     4861 ataccgccag ttgtttaccc tcacaacgtt ccagtaaccg ggcatgttca tcatcagtaa
     4921 cccgtatcgt gagcatcctc tctcgtttca tcggtatcat tacccccatg aacagaaatc
     4981 ccccttacac ggaggcatca gtgaccaaac aggaaaaaac cgcccttaac atggcccgct
     5041 ttatcagaag ccagacatta acgcttctgg agaaactcaa cgagctggac gcggatgaac
     5101 aggcagacat ctgtgaatcg cttcacgacc acgctgatga gctttaccgc agctgcctcg
     5161 cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag
     5221 cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg
     5281 gcgggtgtcg gggcgcagcc atgacccagt cacgtagcga tagcggagtg tatactggct
     5341 taactatgcg gcatcagagc agattgtact gagagtgcac catatatgcg gtgtgaaata
     5401 ccgcacagat gcgtaaggag aaaataccgc atcaggcgct cttccgcttc ctcgctcact
     5461 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     5521 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     5581 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     5641 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     5701 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     5761 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     5821 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     5881 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     5941 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     6001 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     6061 aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     6121 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     6181 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     6241 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgaacaa taaaactgtc
     6301 tgcttacata aacagtaata caaggggtgt tatgagccat attcaacggg aaacgtcttg
     6361 ctctaggccg cgattaaatt ccaacatgga tgctgattta tatgggtata aatgggctcg
     6421 cgataatgtc gggcaatcag gtgcgacaat ctatcgattg tatgggaagc ccgatgcgcc
     6481 agagttgttt ctgaaacatg gcaaaggtag cgttgccaat gatgttacag atgagatggt
     6541 cagactaaac tggctgacgg aatttatgcc tcttccgacc atcaagcatt ttatccgtac
     6601 tcctgatgat gcatggttac tcaccactgc gatccccggg aaaacagcat tccaggtatt
     6661 agaagaatat cctgattcag gtgaaaatat tgttgatgcg ctggcagtgt tcctgcgccg
     6721 gttgcattcg attcctgttt gtaattgtcc ttttaacagc gatcgcgtat ttcgtctcgc
     6781 tcaggcgcaa tcacgaatga ataacggttt ggttgatgcg agtgattttg atgacgagcg
     6841 taatggctgg cctgttgaac aagtctggaa agaaatgcat aaacttttgc cattctcacc
     6901 ggattcagtc gtcactcatg gtgatttctc acttgataac cttatttttg acgaggggaa
     6961 attaataggt tgtattgatg ttggacgagt cggaatcgca gaccgatacc aggatcttgc
     7021 catcctatgg aactgcctcg gtgagttttc tccttcatta cagaaacggc tttttcaaaa
     7081 atatggtatt gataatcctg atatgaataa attgcagttt catttgatgc tcgatgagtt
     7141 tttctaagaa ttaattcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     7201 taggggttcc gcgcacattt ccccgaaaag tgccacctga aattgtaaac gttaatattt
     7261 tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa
     7321 tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag
     7381 tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg
     7441 tctatcaggg cgatggccca ctacgtgaac catcacccta atcaagtttt ttggggtcga
     7501 ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg
     7561 gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg
     7621 cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc
     7681 cgctacaggg cgcgtcccat tcgcca
//
