LOCUS       sequence-337769-        6246 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses pET-15b(+) Mortierella verticillate (delta prenylation 
            motif).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6246)
  TITLE     Evolution of PRL
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6246)
  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            "Unpublished"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: pET-15b(+) Mortierella verticillate (delta
            pre...
FEATURES             Location/Qualifiers
     source          1..6246
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    1..25
                     /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)."
     RBS             40..62
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             81..98
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             108..125
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     primer_bind     complement(725..743)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      739..786
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(961..966,967..983,984..989))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(1016..1034)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1102..1206
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1207..1275,1276..2067)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(1425..1444)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2238..2826
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2727..2746
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2980..2997
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3012..3154
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3080..3099)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3240..3262
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3256..3447)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4759..5841)
                     /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     5803..5822
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5842..5919)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     6125..6144
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     join(6228..6246,1)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6228..6246
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 ggaattgtga gcggataaca attcccctct agaaataatt ttgtttaact ttaagaagga
       61 gatataccat gggcagcagc catcatcatc atcatcacag cagcggcctg gtgccgcgcg
      121 gcagccatat ggcgcaggcg ccgatctctt ctaccacccc gtcccgtatc cacggttctg
      181 aaccgcgtct gctgaaccgt ccggcgctga tcgaatacaa acacctgcgt ttcctggttt
      241 ctgacgcgcc gtctgactct aacctgccgc tgtacgttgc ggagttcgaa cgtcacaccg
      301 ttaaagacgt tgttcgtgtg tgcgacccga cctacggcac cggtctgatg aaagaacgtg
      361 gtatccaggt ttacgattgg ccgttcaacg acggtgaagg tccgccgacc aacatcacca
      421 acgactggct gcgtctggtt ggtctgcgtt tcggtactga cccggaacag atcccgcagg
      481 gtgctatcgc tgttcactgc gttgcgggcc tgggtcgtgc gccgctgctg gttgcgatcg
      541 cgctgatcga agcgggtatg tctccggaag aatctgtgga atttgttcgt accaaacgtc
      601 gtggtgcgct gaacaccaaa caggttaaat acgttcgtga atacaaacgt aaatctcgta
      661 aacgttctct gtaaggatcc ggctgctaac aaagcccgaa aggaagctga gttggctgct
      721 gccaccgctg agcaataact agcataaccc cttggggcct ctaaacgggt cttgaggggt
      781 tttttgctga aaggaggaac tatatccgga tatcccgcaa gaggcccggc agtaccggca
      841 taaccaagcc tatgcctaca gcatccaggg tgacggtgcc gaggatgacg atgagcgcat
      901 tgttagattt catacacggt gcctgactgc gttagcaatt taactgtgat aaactaccgc
      961 attaaagctt atcgatgata agctgtcaaa catgagaatt cttgaagacg aaagggcctc
     1021 gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt
     1081 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
     1141 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
     1201 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc
     1261 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
     1321 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt
     1381 cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta
     1441 ttatcccgtg ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat
     1501 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga
     1561 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca
     1621 acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact
     1681 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc
     1741 acgatgcctg cagcaatggc aacaacgttg cgcaaactat taactggcga actacttact
     1801 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt
     1861 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt
     1921 gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt
     1981 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
     2041 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag
     2101 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     2161 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     2221 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     2281 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     2341 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg
     2401 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     2461 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     2521 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     2581 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     2641 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     2701 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     2761 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     2821 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     2881 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag
     2941 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa
     3001 gcggaagagc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc
     3061 atatatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc cagtatacac
     3121 tccgctatcg ctacgtgact gggtcatggc tgcgccccga cacccgccaa cacccgctga
     3181 cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc
     3241 cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga ggcagctgcg
     3301 gtaaagctca tcagcgtggt cgtgaagcga ttcacagatg tctgcctgtt catccgcgtc
     3361 cagctcgttg agtttctcca gaagcgttaa tgtctggctt ctgataaagc gggccatgtt
     3421 aagggcggtt ttttcctgtt tggtcactga tgcctccgtg taagggggat ttctgttcat
     3481 gggggtaatg ataccgatga aacgagagag gatgctcacg atacgggtta ctgatgatga
     3541 acatgcccgg ttactggaac gttgtgaggg taaacaactg gcggtatgga tgcggcggga
     3601 ccagagaaaa atcactcagg gtcaatgcca gcgcttcgtt aatacagatg taggtgttcc
     3661 acagggtagc cagcagcatc ctgcgatgca gatccggaac ataatggtgc agggcgctga
     3721 cttccgcgtt tccagacttt acgaaacacg gaaaccgaag accattcatg ttgttgctca
     3781 ggtcgcagac gttttgcagc agcagtcgct tcacgttcgc tcgcgtatcg gtgattcatt
     3841 ctgctaacca gtaaggcaac cccgccagcc tagccgggtc ctcaacgaca ggagcacgat
     3901 catgcgcacc cgtggccagg acccaacgct gcccgagatg cgccgcgtgc ggctgctgga
     3961 gatggcggac gcgatggata tgttctgcca agggttggtt tgcgcattca cagttctccg
     4021 caagaattga ttggctccaa ttcttggagt ggtgaatccg ttagcgaggt gccgccggct
     4081 tccattcagg tcgaggtggc ccggctccat gcaccgcgac gcaacgcggg gaggcagaca
     4141 aggtataggg cggcgcctac aatccatgcc aacccgttcc atgtgctcgc cgaggcggca
     4201 taaatcgccg tgacgatcag cggtccagtg atcgaagtta ggctggtaag agccgcgagc
     4261 gatccttgaa gctgtccctg atggtcgtca tctacctgcc tggacagcat ggcctgcaac
     4321 gcgggcatcc cgatgccgcc ggaagcgaga agaatcataa tggggaaggc catccagcct
     4381 cgcgtcgcga acgccagcaa gacgtagccc agcgcgtcgg ccgccatgcc ggcgataatg
     4441 gcctgcttct cgccgaaacg tttggtggcg ggaccagtga cgaaggcttg agcgagggcg
     4501 tgcaagattc cgaataccgc aagcgacagg ccgatcatcg tcgcgctcca gcgaaagcgg
     4561 tcctcgccga aaatgaccca gagcgctgcc ggcacctgtc ctacgagttg catgataaag
     4621 aagacagtca taagtgcggc gacgatagtc atgccccgcg cccaccggaa ggagctgact
     4681 gggttgaagg ctctcaaggg catcggtcga gatcccggtg cctaatgagt gagctaactt
     4741 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
     4801 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ccagggtggt
     4861 ttttcttttc accagtgaga cgggcaacag ctgattgccc ttcaccgcct ggccctgaga
     4921 gagttgcagc aagcggtcca cgctggtttg ccccagcagg cgaaaatcct gtttgatggt
     4981 ggttaacggc gggatataac atgagctgtc ttcggtatcg tcgtatccca ctaccgagat
     5041 atccgcacca acgcgcagcc cggactcggt aatggcgcgc attgcgccca gcgccatctg
     5101 atcgttggca accagcatcg cagtgggaac gatgccctca ttcagcattt gcatggtttg
     5161 ttgaaaaccg gacatggcac tccagtcgcc ttcccgttcc gctatcggct gaatttgatt
     5221 gcgagtgaga tatttatgcc agccagccag acgcagacgc gccgagacag aacttaatgg
     5281 gcccgctaac agcgcgattt gctggtgacc caatgcgacc agatgctcca cgcccagtcg
     5341 cgtaccgtct tcatgggaga aaataatact gttgatgggt gtctggtcag agacatcaag
     5401 aaataacgcc ggaacattag tgcaggcagc ttccacagca atggcatcct ggtcatccag
     5461 cggatagtta atgatcagcc cactgacgcg ttgcgcgaga agattgtgca ccgccgcttt
     5521 acaggcttcg acgccgcttc gttctaccat cgacaccacc acgctggcac ccagttgatc
     5581 ggcgcgagat ttaatcgccg cgacaatttg cgacggcgcg tgcagggcca gactggaggt
     5641 ggcaacgcca atcagcaacg actgtttgcc cgccagttgt tgtgccacgc ggttgggaat
     5701 gtaattcagc tccgccatcg ccgcttccac tttttcccgc gttttcgcag aaacgtggct
     5761 ggcctggttc accacgcggg aaacggtctg ataagagaca ccggcatact ctgcgacatc
     5821 gtataacgtt actggtttca cattcaccac cctgaattga ctctcttccg ggcgctatca
     5881 tgccataccg cgaaaggttt tgcgccattc gatggtgtcc gggatctcga cgctctccct
     5941 tatgcgactc ctgcattagg aagcagccca gtagtaggtt gaggccgttg agcaccgccg
     6001 ccgcaaggaa tggtgcatgc aaggagatgg cgcccaacag tcccccggcc acggggcctg
     6061 ccaccatacc cacgccgaaa caagcgctca tgagcccgaa gtggcgagcc cgatcttccc
     6121 catcggtgat gtcggcgata taggcgccag caaccgcacc tgtggcgccg gtgatgccgg
     6181 ccacgatgcg tccggcgtag aggatcgaga tctcgatccc gcgaaattaa tacgactcac
     6241 tatagg
//
