LOCUS       sequence-368108-        5156 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Pom152 N-terminus 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 5156)
  AUTHORS   Ken Belanger Lab
  TITLE     Belanger lab Plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5156)
  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            "Unpublished"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: KBB609
FEATURES             Location/Qualifiers
     source          1..5156
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     123..139
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     175..194
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        175..193
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    194..218
                     /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             222..244
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             255..272
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             273..293
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(1015..1033)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1029..1075
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        1321..1398
                     /gene="lacI"
                     /label=lacI promoter
     CDS             1399..2481
                     /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(1418..1437)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(2530..2535,2536..2553,2554..2560)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2568..2584
                     /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     2573..2595
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2592..2608
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2592..2608
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2624..2640)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      complement(2932..3520)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3012..3031)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3691..4482,4483..4551))
                     /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     4314..4333
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4552..4656)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4683..5138
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4770..4789)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4980..5001
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa
       61 agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg
      121 ttgtaaaacg acggccagtg ccgaattctg ctggactttc caaaatgtcg agcttaatac
      181 gactcactat aggggaattg tgagcggata acaattcccc ttttgtttaa ctttaagaag
      241 gagaggccac catgcatcac catcaccatc acgaaaacct gtattttcag ggcgcgatcg
      301 ccattatgac cacatggaga aaactttata cgaaagaatt aagtgtaaca ggttcagcca
      361 ttaaccatca tcgtattttt gattcttccg ctcattttaa gggtgctcta acaatcaaga
      421 tcctacctga aaacacagct atgtttaatc ctcttcatga atcatattgt ttgcccatgg
      481 ataccaatct ttttaaaatt aactcaattg acgtacccat cagaataaac tcaactgaag
      541 agattgaata tatcgagttg gaatatagag atctttacac aaattcggta gagttacggt
      601 ctttgagtaa gaaagacttc aaaatcatcg ataaccccaa atccttttta aaaaaagatc
      661 agtcagttct aaagtcacat tcaaacgatt ttgaagaagg cagtacaatt cgttatttgg
      721 ctgtaactct tcaagatatt ggcttttatc aaatcaaaaa aattgttgat tcaaaaaaac
      781 tgaatttaaa gatccatcag tcacacttgg ttgtacccta ctgtccaatt gcatctatta
      841 ctggaactgg aagcaatgat agatgtattg gcgattcaga taatgtttct tttgaaattc
      901 aaggtgtgcc cccaatgaaa ttagcatact ctaagatcgt taatggttaa acgcgttaaa
      961 cgcggccgct cgaggtttaa acggccggcc acatgactga ctgagccacg ataaccgctg
     1021 agcaataact agcataaccc cttggggcct ctaaacgggt cttgaggggt tttttcccac
     1081 cagccttgtc ctaatccgcg gcggttcctg gccttttgct ggccttttgc tcacatgttc
     1141 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
     1201 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag
     1261 cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg cataaattcc
     1321 gacaccatcg aatggcgcaa aacctttcgc ggtatggcat gatagcgccc ggaagagagt
     1381 caattcaggg tggtgaatgt gaaaccagta acgttatacg atgtcgcaga gtatgccggt
     1441 gtctcttatc agaccgtttc ccgcgtggtg aaccaggcca gccacgtttc tgcgaaaacg
     1501 cgggaaaaag tggaagcggc gatggcggag ctgaattaca ttcccaaccg cgtggcacaa
     1561 caactggcgg gcaaacagtc gttgctgatt ggcgttgcca cctccagtct ggccctgcac
     1621 gcgccgtcgc aaattgtcgc ggcgattaaa tctcgcgccg atcaactggg tgccagcgtg
     1681 gtggtgtcga tggtagaacg aagcggcgtc gaagcctgta aagcggcggt gcacaatctt
     1741 ctcgcgcaac gagtcagtgg gctgatcatt aactatccgc tggatgacca ggatgccatt
     1801 gctgtggaag ctgcctgcac taatgttccg gcgttatttc ttgatgtctc tgaccagaca
     1861 cccatcaaca gtattatttt ctcccatgaa gacggtacgc gactgggcgt ggagcatctg
     1921 gtcgcattgg gtcaccagca aatcgcgctg ttagcgggcc cattaagttc tgtctcggcg
     1981 cgtctgcgtc tggctggctg gcataaatat ctcactcgca atcaaattca gccgatagcg
     2041 gaacgggaag gcgactggag tgccatgtcc ggttttcaac aaaccatgca aatgctgaat
     2101 gagggcatcg ttcccactgc gatgctggtt gccaacgatc agatggcgct gggcgcaatg
     2161 cgcgccatta ccgagtccgg gctgcgcgtt ggtgcggata tctcggtagt gggatacgac
     2221 gataccgaag acagctcatg ttatatcccg ccgttaacca ccatcaaaca ggattttcgc
     2281 ctgctggggc aaaccagcgt ggaccgcttg ctgcaactct ctcagggcca ggcggtgaag
     2341 ggcaatcagc tgttgcccgt ctcactggtg aaaagaaaaa ccaccctggc gcccaatacg
     2401 caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc
     2461 cgactggaaa gcgggcagtg agcgcaacgc aattaatgta agttagctca ctcattaggc
     2521 accccaggct ttacacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata
     2581 acaatttcac acaggaaaca gctatgacca tgattacgga ttcactggcc gtcgttttac
     2641 aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc
     2701 ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tggatccgct cttccgcttc
     2761 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     2821 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     2881 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     2941 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     3001 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     3061 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     3121 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     3181 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     3241 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     3301 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     3361 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     3421 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     3481 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     3541 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     3601 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     3661 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     3721 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     3781 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     3841 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     3901 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     3961 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     4021 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     4081 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     4141 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     4201 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     4261 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     4321 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     4381 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     4441 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     4501 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     4561 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     4621 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     4681 tgacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     4741 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     4801 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     4861 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg
     4921 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     4981 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
     5041 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
     5101 taacgcgaat tttaacaaaa tattaacgct tacaatttcc attcgccatt caggct
//
