LOCUS       sequence-368078-        4934 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Pom152 central repeats 4 and 5 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 4934)
  AUTHORS   Ken Belanger Lab
  TITLE     Belanger lab Plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4934)
  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            "Unpublished"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: KBB611
FEATURES             Location/Qualifiers
     source          1..4934
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     80..96
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     132..151
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        132..150
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    151..175
                     /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             179..201
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             212..229
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             230..250
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(750..768)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      764..810
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        1056..1133
                     /gene="lacI"
                     /label=lacI promoter
     CDS             1134..2216
                     /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(1153..1172)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(2265..2270,2271..2288,2289..2295)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2303..2319
                     /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     2308..2330
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2327..2343
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2327..2343
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2359..2375)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      complement(2667..3255)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2747..2766)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3426..4217,4218..4286))
                     /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     4049..4068
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4287..4391)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4418..4873
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4505..4524)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4715..4736
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat taagttgggt aacgccaggg
       61 ttttcccagt cacgacgttg taaaacgacg gccagtgccg aattctgctg gactttccaa
      121 aatgtcgagc ttaatacgac tcactatagg ggaattgtga gcggataaca attccccttt
      181 tgtttaactt taagaaggag aggccaccat gcatcaccat caccatcacg aaaacctgta
      241 ttttcagggc gcgatcgccg aacctatcaa agaagctaaa attaagcacg gttctgtcac
      301 tgaaattccg ctcaaattaa gtggagaggg gccattcacc gttaagttta aacacatgga
      361 ttacgatgga aacattgtga aagaatttga aaacaaattt cagaatagtt ataagcccgc
      421 tttgaaagta agtaaagaag ggctttacca attggtcgat attcgtgact caagttgcca
      481 aggtaatgtg atttaccgga acagtctgta taaggtatct ttcttggaga agccaaaatt
      541 tgcaattcaa gacaatcatc acattacaaa agtaacggaa aatttattct ctaaagaaga
      601 agtctgtcaa gggatggaag gtacggttga tttggctcta tttggttctc caccattcat
      661 attagaatat gatttgatgg cataaacgcg ttaaacgcgg ccgctcgagg tttaaacggc
      721 cggccacatg actgactgag ccacgataac cgctgagcaa taactagcat aaccccttgg
      781 ggcctctaaa cgggtcttga ggggtttttt cccaccagcc ttgtcctaat ccgcggcggt
      841 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
      901 tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg
      961 agcgcagcga gtcagtgagc gaggaagcgg aagagcgcct gatgcggtat tttctcctta
     1021 cgcatctgtg cggtatttca caccgcataa attccgacac catcgaatgg cgcaaaacct
     1081 ttcgcggtat ggcatgatag cgcccggaag agagtcaatt cagggtggtg aatgtgaaac
     1141 cagtaacgtt atacgatgtc gcagagtatg ccggtgtctc ttatcagacc gtttcccgcg
     1201 tggtgaacca ggccagccac gtttctgcga aaacgcggga aaaagtggaa gcggcgatgg
     1261 cggagctgaa ttacattccc aaccgcgtgg cacaacaact ggcgggcaaa cagtcgttgc
     1321 tgattggcgt tgccacctcc agtctggccc tgcacgcgcc gtcgcaaatt gtcgcggcga
     1381 ttaaatctcg cgccgatcaa ctgggtgcca gcgtggtggt gtcgatggta gaacgaagcg
     1441 gcgtcgaagc ctgtaaagcg gcggtgcaca atcttctcgc gcaacgagtc agtgggctga
     1501 tcattaacta tccgctggat gaccaggatg ccattgctgt ggaagctgcc tgcactaatg
     1561 ttccggcgtt atttcttgat gtctctgacc agacacccat caacagtatt attttctccc
     1621 atgaagacgg tacgcgactg ggcgtggagc atctggtcgc attgggtcac cagcaaatcg
     1681 cgctgttagc gggcccatta agttctgtct cggcgcgtct gcgtctggct ggctggcata
     1741 aatatctcac tcgcaatcaa attcagccga tagcggaacg ggaaggcgac tggagtgcca
     1801 tgtccggttt tcaacaaacc atgcaaatgc tgaatgaggg catcgttccc actgcgatgc
     1861 tggttgccaa cgatcagatg gcgctgggcg caatgcgcgc cattaccgag tccgggctgc
     1921 gcgttggtgc ggatatctcg gtagtgggat acgacgatac cgaagacagc tcatgttata
     1981 tcccgccgtt aaccaccatc aaacaggatt ttcgcctgct ggggcaaacc agcgtggacc
     2041 gcttgctgca actctctcag ggccaggcgg tgaagggcaa tcagctgttg cccgtctcac
     2101 tggtgaaaag aaaaaccacc ctggcgccca atacgcaaac cgcctctccc cgcgcgttgg
     2161 ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc
     2221 aacgcaatta atgtaagtta gctcactcat taggcacccc aggctttaca ctttatgctt
     2281 ccggctcgta tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat
     2341 gaccatgatt acggattcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg
     2401 cgttacccaa cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga
     2461 agaggcccgc accgatggat ccgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     2521 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     2581 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     2641 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     2701 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     2761 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     2821 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     2881 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     2941 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     3001 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     3061 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     3121 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     3181 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     3241 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     3301 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     3361 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     3421 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     3481 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     3541 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     3601 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     3661 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     3721 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     3781 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     3841 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     3901 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     3961 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     4021 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     4081 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     4141 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     4201 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     4261 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     4321 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     4381 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg cgccctgtag cggcgcatta
     4441 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
     4501 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     4561 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     4621 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     4681 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     4741 acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc
     4801 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta
     4861 acgcttacaa tttccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc
     4921 gggcctcttc gcta
//
