LOCUS       sequence-274534-        6242 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of 4'-phosphopantetheinyl transferase gene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6242)
  TITLE     NEB Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6242)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pBAD-MBP-His-SFP
FEATURES             Location/Qualifiers
     source          1..6242
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..285
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     208..227
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             319..1422
                     /codon_start=1
                     /gene="malE (mutated)"
                     /product="maltose binding protein from E. coli"
                     /label=MBP
                     /note="This version of the gene does not encode a signal 
                     sequence, so MBP will remain in the cytosol."
                     /translation="MVKIEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKL
                     EEKFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNGK
                     LIAYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWPLI
                     AADGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAFNKG
                     ETAMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKELAKEF
                     LENYLLTDEGLEAVNKDKPLGAVALKSYEEELAKDPRIAATMENAQKGEIMPNIPQMSA
                     FWYAVRTAVINAASGRQTVDEALKDAQT"
     primer_bind     1396..1419
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     CDS             1435..1464
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             1438..1467
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             1441..1470
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             1444..1473
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             1447..1476
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             1450..1479
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             1489..1512
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             2203..2571
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="AKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDDV
                     TLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGRE
                     FALRDPAGNCVHFVAEEQD"
     CDS             2572..2601
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     primer_bind     complement(2643..2660)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      2813..2899
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2991..3018
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        3037..3128
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3129..3197,3198..3989)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(3347..3366)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4160..4748
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4649..4668
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4902..4919
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4934..5074
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(5000..5019)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5160..5182
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(5338..6216)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
ORIGIN
        1 aagaaaccaa ttgtccatat tgcatcagac attgccgtca ctgcgtcttt tactggctct
       61 tctcgctaac caaaccggta accccgctta ttaaaagcat tctgtaacaa agcgggacca
      121 aagccatgac aaaaacgcgt aacaaaagtg tctataatca cggcagaaaa gtccacattg
      181 attatttgca cggcgtcaca ctttgctatg ccatagcatt tttatccata agattagcgg
      241 atcctacctg acgcttttta tcgcaactct ctactgtttc tccatacccg ttttttgggc
      301 taacaggagg aattaaccat ggtaaaaatc gaagaaggta aactggtaat ctggattaac
      361 ggcgataaag gctataacgg tctcgctgaa gtcggtaaga aattcgagaa agataccgga
      421 attaaagtca ccgttgagca tccggataaa ctggaagaga aattcccaca ggttgcggca
      481 actggcgatg gccctgacat tatcttctgg gcacacgacc gctttggtgg ctacgctcaa
      541 tctggcctgt tggctgaaat caccccggac aaagcgttcc aggacaagct gtatccgttt
      601 acctgggatg ccgtacgtta caacggcaag ctgattgctt acccgatcgc tgttgaagcg
      661 ttatcgctga tttataacaa agatctgctg ccgaacccgc caaaaacctg ggaagagatc
      721 ccggcgctgg ataaagaact gaaagcgaaa ggtaagagcg cgctgatgtt caacctgcaa
      781 gaaccgtact tcacctggcc gctgattgct gctgacgggg gttatgcgtt caagtatgaa
      841 aacggcaagt acgacattaa agacgtgggc gtggataacg ctggcgcgaa agcgggtctg
      901 accttcctgg ttgacctgat taaaaacaaa cacatgaatg cagacaccga ttactccatc
      961 gcagaagctg cctttaataa aggcgaaaca gcgatgacca tcaacggccc gtgggcatgg
     1021 tccaacatcg acaccagcaa agtgaattat ggtgtaacgg tactgccgac cttcaagggt
     1081 caaccatcca aaccgttcgt tggcgtgctg agcgcaggta ttaacgccgc cagtccgaac
     1141 aaagagctgg caaaagagtt cctcgaaaac tatctgctga ctgatgaagg tctggaagcg
     1201 gttaataaag acaaaccgct gggtgccgta gcgctgaagt cttacgagga agagttggcg
     1261 aaagatccac gtattgccgc cactatggaa aacgcccaga aaggtgaaat catgccgaac
     1321 atcccgcaga tgtccgcttt ctggtatgcc gtgcgtactg cggtgatcaa cgccgccagc
     1381 ggtcgtcaga ctgtcgatga agccctgaaa gacgcgcaga ctaattcgag ctcgcatcat
     1441 catcatcatc atcatcatca tcatcatcat catcatcatc ctgcaggctt agaggttctg
     1501 ttccaggggc ctatgaagat ttacggaatt tatatggacc gcccgctttc acaggaagaa
     1561 aatgaacggt tcatgacttt catatcacct gaaaaacggg agaaatgccg gagattttat
     1621 cataaagaag atgctcaccg caccctgctg ggagatgtgc tcgttcgctc agtcataagc
     1681 aggcagtatc agttggacaa atccgatatc cgctttagca cgcaggaata cgggaagccg
     1741 tgcatccctg atcttcccga cgctcatttc aacatttctc actccggccg ctgggtcatt
     1801 ggtgcgtttg attcacagcc gatcggcata gatatcgaaa aaacgaaacc gatcagcctt
     1861 gagatcgcca agcgcttctt ttcaaaaaca gagtacagcg accttttagc aaaagacaag
     1921 gacgagcaga cagactattt ttatcatcta tggtcaatga aagaaagctt tatcaaacag
     1981 gaaggcaaag gcttatcgct tccgcttgat tccttttcag tgcgcctgca tcaggacgga
     2041 caagtatcca ttgagcttcc ggacagccat tccccatgct atatcaaaac gtatgaggtc
     2101 gatcccggct acaaaatggc tgtatgcgcc gcacaccctg atttccccga ggatatcaca
     2161 atggtctcgt acgaagagct tttataagga tccggcgcgc cggccaagtt gaccagtgcc
     2221 gttccggtgc tcaccgcgcg cgacgtcgcc ggagcggtcg agttctggac cgaccggctc
     2281 gggttctccc gggacttcgt ggaggacgac ttcgccggtg tggtccggga cgacgtgacc
     2341 ctgttcatca gcgcggtcca ggaccaggtg gtgccggaca acaccctggc ctgggtgtgg
     2401 gtgcgcggcc tggacgagct gtacgccgag tggtcggagg tcgtgtccac gaacttccgg
     2461 gacgcctccg ggccggccat gaccgagatc ggcgagcagc cgtgggggcg ggagttcgcc
     2521 ctgcgcgacc cggccggcaa ctgcgtgcac ttcgtggccg aggagcagga ccatcatcat
     2581 catcatcatc atcatcatca ttaaaagctt ggctgttttg gcggatgaga gaagattttc
     2641 agcctgatac agattaaatc agaacgcaga agcggtctga taaaacagaa tttgcctggc
     2701 ggcagtagcg cggtggtccc acctgacccc atgccgaact cagaagtgaa acgccgtagc
     2761 gccgatggta gtgtggggtc tccccatgcg agagtaggga actgccaggc atcaaataaa
     2821 acgaaaggct cagtcgaaag actgggcctt tcgttttatc tgttgtttgt cggtgaacgc
     2881 tctcctgagt aggacaaatc cgccgggagc ggatttgaac gttgcgaagc aacggcccgg
     2941 agggtggcgg gcaggacgcc cgccataaac tgccaggcat caaattaagc agaaggccat
     3001 cctgacggat ggcctttttg cgtttctaca aactcttttg tttatttttc taaatacatt
     3061 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     3121 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
     3181 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
     3241 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
     3301 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
     3361 tattatcccg tgttgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
     3421 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
     3481 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
     3541 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
     3601 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
     3661 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
     3721 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
     3781 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
     3841 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
     3901 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
     3961 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
     4021 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     4081 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     4141 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     4201 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     4261 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc
     4321 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     4381 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     4441 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     4501 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     4561 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     4621 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     4681 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     4741 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     4801 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg
     4861 agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg
     4921 aagcggaaga gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc
     4981 gcatatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc cagtatacac
     5041 tccgctatcg ctacgtgact gggtcatggc tgcgccccga cacccgccaa cacccgctga
     5101 cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc
     5161 cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga ggcagcagat
     5221 caattcgcgc gcgaaggcga agcggcatgc ataatgtgcc tgtcaaatgg acgaagcagg
     5281 gattctgcaa accctatgct actccgtcaa gccgtcaatt gtctgattcg ttaccaatta
     5341 tgacaacttg acggctacat cattcacttt ttcttcacaa ccggcacgga actcgctcgg
     5401 gctggccccg gtgcattttt taaatacccg cgagaaatag agttgatcgt caaaaccaac
     5461 attgcgaccg acggtggcga taggcatccg ggtggtgctc aaaagcagct tcgcctggct
     5521 gatacgttgg tcctcgcgcc agcttaagac gctaatccct aactgctggc ggaaaagatg
     5581 tgacagacgc gacggcgaca agcaaacatg ctgtgcgacg ctggcgatat caaaattgct
     5641 gtctgccagg tgatcgctga tgtactgaca agcctcgcgt acccgattat ccatcggtgg
     5701 atggagcgac tcgttaatcg cttccatgcg ccgcagtaac aattgctcaa gcagatttat
     5761 cgccagcagc tccgaatagc gcccttcccc ttgcccggcg ttaatgattt gcccaaacag
     5821 gtcgctgaaa tgcggctggt gcgcttcatc cgggcgaaag aaccccgtat tggcaaatat
     5881 tgacggccag ttaagccatt catgccagta ggcgcgcgga cgaaagtaaa cccactggtg
     5941 ataccattcg cgagcctccg gatgacgacc gtagtgatga atctctcctg gcgggaacag
     6001 caaaatatca cccggtcggc aaacaaattc tcgtccctga tttttcacca ccccctgacc
     6061 gcgaatggtg agattgagaa tataaccttt cattcccagc ggtcggtcga taaaaaaatc
     6121 gagataaccg ttggcctcaa tcggcgttaa acccgccacc agatgggcat taaacgagta
     6181 tcccggcagc aggggatcat tttgcgcttc agccatactt ttcatactcc cgccattcag
     6241 ag
//
