LOCUS       sequence-368105-        4580 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Pom152 full repeat domain in pEX-N-His for bacterial expression and 
            purification.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4580)
  AUTHORS   Ken Belanger Lab
  TITLE     Belanger lab Plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4580)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: KBB610
FEATURES             Location/Qualifiers
     source          1..4580
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     28..47
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        28..46
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    47..71
                     /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             75..97
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             108..125
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             126..146
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(292..310)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      306..352
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        598..675
                     /gene="lacI"
                     /label=lacI promoter
     CDS             676..1758
                     /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(695..714)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(1807..1812,1813..1830,1831..1837)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1845..1861
                     /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     1850..1872
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1869..1885
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1869..1885
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1901..1917)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      complement(2209..2797)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2289..2308)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2968..3759,3760..3828))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     3591..3610
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3829..3933)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      3960..4415
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4047..4066)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4257..4278
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4556..4572
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 ctgctggact ttccaaaatg tcgagcttaa tacgactcac tataggggaa ttgtgagcgg
       61 ataacaattc cccttttgtt taactttaag aaggagaggc caccatgcat caccatcacc
      121 atcacgaaaa cctgtatttt cagggcgcga tcgccgtctg tcaagggatg gaaggtacgg
      181 ttgatttggc tctatttggt tctccaccat tcatattaga atattaaacg cgttaaacgc
      241 ggccgctcga ggtttaaacg gccggccaca tgactgactg agccacgata accgctgagc
      301 aataactagc ataacccctt ggggcctcta aacgggtctt gaggggtttt ttcccaccag
      361 ccttgtccta atccgcggcg gttcctggcc ttttgctggc cttttgctca catgttcttt
      421 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
      481 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc
      541 ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat aaattccgac
      601 accatcgaat ggcgcaaaac ctttcgcggt atggcatgat agcgcccgga agagagtcaa
      661 ttcagggtgg tgaatgtgaa accagtaacg ttatacgatg tcgcagagta tgccggtgtc
      721 tcttatcaga ccgtttcccg cgtggtgaac caggccagcc acgtttctgc gaaaacgcgg
      781 gaaaaagtgg aagcggcgat ggcggagctg aattacattc ccaaccgcgt ggcacaacaa
      841 ctggcgggca aacagtcgtt gctgattggc gttgccacct ccagtctggc cctgcacgcg
      901 ccgtcgcaaa ttgtcgcggc gattaaatct cgcgccgatc aactgggtgc cagcgtggtg
      961 gtgtcgatgg tagaacgaag cggcgtcgaa gcctgtaaag cggcggtgca caatcttctc
     1021 gcgcaacgag tcagtgggct gatcattaac tatccgctgg atgaccagga tgccattgct
     1081 gtggaagctg cctgcactaa tgttccggcg ttatttcttg atgtctctga ccagacaccc
     1141 atcaacagta ttattttctc ccatgaagac ggtacgcgac tgggcgtgga gcatctggtc
     1201 gcattgggtc accagcaaat cgcgctgtta gcgggcccat taagttctgt ctcggcgcgt
     1261 ctgcgtctgg ctggctggca taaatatctc actcgcaatc aaattcagcc gatagcggaa
     1321 cgggaaggcg actggagtgc catgtccggt tttcaacaaa ccatgcaaat gctgaatgag
     1381 ggcatcgttc ccactgcgat gctggttgcc aacgatcaga tggcgctggg cgcaatgcgc
     1441 gccattaccg agtccgggct gcgcgttggt gcggatatct cggtagtggg atacgacgat
     1501 accgaagaca gctcatgtta tatcccgccg ttaaccacca tcaaacagga ttttcgcctg
     1561 ctggggcaaa ccagcgtgga ccgcttgctg caactctctc agggccaggc ggtgaagggc
     1621 aatcagctgt tgcccgtctc actggtgaaa agaaaaacca ccctggcgcc caatacgcaa
     1681 accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga
     1741 ctggaaagcg ggcagtgagc gcaacgcaat taatgtaagt tagctcactc attaggcacc
     1801 ccaggcttta cactttatgc ttccggctcg tatgttgtgt ggaattgtga gcggataaca
     1861 atttcacaca ggaaacagct atgaccatga ttacggattc actggccgtc gttttacaac
     1921 gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca catccccctt
     1981 tcgccagctg gcgtaatagc gaagaggccc gcaccgatgg atccgctctt ccgcttcctc
     2041 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     2101 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     2161 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     2221 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     2281 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     2341 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     2401 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     2461 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     2521 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     2581 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     2641 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     2701 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     2761 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     2821 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     2881 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     2941 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     3001 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     3061 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     3121 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     3181 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     3241 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     3301 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     3361 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     3421 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     3481 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     3541 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     3601 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     3661 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     3721 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     3781 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     3841 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     3901 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
     3961 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     4021 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     4081 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     4141 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
     4201 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     4261 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
     4321 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     4381 cgcgaatttt aacaaaatat taacgcttac aatttccatt cgccattcag gctgcgcaac
     4441 tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga
     4501 tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa
     4561 acgacggcca gtgccgaatt
//
