LOCUS       sequence-469255-        6616 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  over-express E. coli holB (Pol III delta prime subunit).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6616)
  AUTHORS   O'Donnell Lab
  TITLE     O'Donnell Lab Batch 1
  JOURNAL   unpublished
REFERENCE   2  (bases 1 to 6616)
  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            "unpublished"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: Ec holB-pET(11a)(ACH-)
FEATURES             Location/Qualifiers
     source          1..6616
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(643..1434,1435..1503))
                     /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     1266..1285
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1504..1608)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1672..1690
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(1920..1967)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     1963..1981
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     protein_bind    3058..3082
                     /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     complement(3082..3101)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3083..3101)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3185..3204)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        3410..3487
                     /gene="lacI"
                     /label=lacI promoter
     CDS             3488..4570
                     /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(3507..3526)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             5882..6073
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(6067..6089)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    6175..6314
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     6230..6249
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(6329..6346)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(6500..6616,1..472))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6580..6599)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
       61 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
      121 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
      181 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
      241 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
      301 gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt
      361 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
      421 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
      481 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
      541 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
      601 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
      661 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
      721 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
      781 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
      841 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
      901 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctgc
      961 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     1021 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     1081 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     1141 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     1201 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaac
     1261 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     1321 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     1381 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     1441 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     1501 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     1561 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     1621 aaaagtgcca cctgctaaga aaccattatt atcatgacat taacctataa aaataggcgt
     1681 atcacgaggc cctttcgtct tcaagaattc tcatgtttga cagcttatca tcgaccagct
     1741 ttaatgcggt agtttatcac agttaaattg ctaacgcagt caggcaccgt gtatgaaatc
     1801 taacaatgcg ctcatcgtca tcctcggcac cgtcaccctg gatgctgtag gcataggctt
     1861 ggttatgccg gtactgccgg gcctcttgcg ggatatccgg atatagttcc tcctttcagc
     1921 aaaaaacccc tcaagacccg tttagaggcc ccaaggggtt atgctagtta ttgctcagcg
     1981 gtggcagcag ccaactcagc ttcctttcgg gctttgttag cagccggatc cttaaagatg
     2041 aggaaccggt agcacaacgc ccggttgcag gtaatgctca atacgcagta aaagatcggt
     2101 gatgagaagc tcgcggttga tgcctgtaac agacattaac tgttcacgaa tgtggcaaac
     2161 atcccccagt atagcctgca ggcgcgaggg agaaagatgg tttgccagtt cggcgaccag
     2221 gcccggcaca tcaacattgg tcacctgcgc agcaccatga tggcgtttta gcgcatccat
     2281 cagcaacgtt gccagccagt gtaaacgcgc cggagcttgt tcatgattaa gggccgctag
     2341 cagcgaatac cagtcgcccg atggcacgct atatgccaac gcctgacaca atgtttcacg
     2401 agcctgccag ttatctccct gaaacaacgc cagtgccgcg ccaggcgaac cggcgcttaa
     2461 gcgcaatgcg gcaagtaatg catcctgtga cattgtcact tcgcgtgaaa gccaggtcac
     2521 ggcgtactgt tccggcggcg gcgcaaggta atgtaaccga caacgactac gtaatgttgc
     2581 cagtaaacgt tcaggctcgc gggtagccag gaaaaaccaa gtttctgctg gtggctcttc
     2641 aagcgttttc agcaatgcgt tagccgcggc gtcggttagt aaggcagcat cggttaccca
     2701 aacgactttc gcaccaccta agcgtgcgtg ctcattcagc ttttcggtga cctcacgtac
     2761 cgcatcaacg cccagcgtat tttttccttt ttcgggagcc agggtgtagt aatcgggatg
     2821 cgtgccagcc tgcatcaact gacatccacg acagtgaccg caacttttgt ggccctgcgg
     2881 ttgttggcag agtaaataac ggctcagggc gtagattaaa gcatcatcgc ccatgcccgg
     2941 taacgcctga atgagtagcg catggtgacc tcttccggcc tgatagctgg ctaccagttt
     3001 ttcgaaatca ggtcgtaacc atggatacca tctcatattt atatctcctt ctagagggga
     3061 attgttatcc gctcacaatt cccctatagt gagtcgtatt aatttcgcgg gatcgagatc
     3121 tcgatcctct acgccggacg catcgtggcc ggcatcaccg gcgccacagg tgcggttgct
     3181 ggcgcctata tcgccgacat caccgatggg gaagatcggg ctcgccactt cgggctcatg
     3241 agcgcttgtt tcggcgtggg tatggtggca ggccccgtgg ccgggggact gttgggcgcc
     3301 atctccttgc atgcaccatt ccttgcggcg gcggtgctca acggcctcaa cctactactg
     3361 ggctgcttcc taatgcagga gtcgcataag ggagagcgtc gagatcccgg acaccatcga
     3421 atggcgcaaa acctttcgcg gtatggcatg atagcgcccg gaagagagtc aattcagggt
     3481 ggtgaatgtg aaaccagtaa cgttatacga tgtcgcagag tatgccggtg tctcttatca
     3541 gaccgtttcc cgcgtggtga accaggccag ccacgtttct gcgaaaacgc gggaaaaagt
     3601 ggaagcggcg atggcggagc tgaattacat tcccaaccgc gtggcacaac aactggcggg
     3661 caaacagtcg ttgctgattg gcgttgccac ctccagtctg gccctgcacg cgccgtcgca
     3721 aattgtcgcg gcgattaaat ctcgcgccga tcaactgggt gccagcgtgg tggtgtcgat
     3781 ggtagaacga agcggcgtcg aagcctgtaa agcggcggtg cacaatcttc tcgcgcaacg
     3841 cgtcagtggg ctgatcatta actatccgct ggatgaccag gatgccattg ctgtggaagc
     3901 tgcctgcact aatgttccgg cgttatttct tgatgtctct gaccagacac ccatcaacag
     3961 tattattttc tcccatgaag acggtacgcg actgggcgtg gagcatctgg tcgcattggg
     4021 tcaccagcaa atcgcgctgt tagcgggccc attaagttct gtctcggcgc gtctgcgtct
     4081 ggctggctgg cataaatatc tcactcgcaa tcaaattcag ccgatagcgg aacgggaagg
     4141 cgactggagt gccatgtccg gttttcaaca aaccatgcaa atgctgaatg agggcatcgt
     4201 tcccactgcg atgctggttg ccaacgatca gatggcgctg ggcgcaatgc gcgccattac
     4261 cgagtccggg ctgcgcgttg gtgcggatat ctcggtagtg ggatacgacg ataccgaaga
     4321 cagctcatgt tatatcccgc cgttaaccac catcaaacag gattttcgcc tgctggggca
     4381 aaccagcgtg gaccgcttgc tgcaactctc tcagggccag gcggtgaagg gcaatcagct
     4441 gttgcccgtc tcactggtga aaagaaaaac caccctggcg cccaatacgc aaaccgcctc
     4501 tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag
     4561 cgggcagtga gcgcaacgca attaatgtaa gttagctcac tcattaggca ccgggatctc
     4621 gaccgatgcc cttgagagcc ttcaacccag tcagctcctt ccggtgggcg cggggcatga
     4681 ctatcgtcgc cgcacttatg actgtcttct ttatcatgca actcgtagga caggtgccgg
     4741 cagcgctctg ggtcattttc ggcgaggacc gctttcgctg gagcgcgacg atgatcggcc
     4801 tgtcgcttgc ggtattcgga atcttgcacg ccctcgctca agccttcgtc actggtcccg
     4861 ccaccaaacg tttcggcgag aagcaggcca ttatcgccgg catggcggcc gacgcgctgg
     4921 gctacgtctt gctggcgttc gcgacgcgag gctggatggc cttccccatt atgattcttc
     4981 tcgcttccgg cggcatcggg atgcccgcgt tgcaggccat gctgtccagg caggtagatg
     5041 acgaccatca gggacagctt caaggatcgc tcgcggctct taccagccta acttcgatca
     5101 ttggaccgct gatcgtcacg gcgatttatg ccgcctcggc gagcacatgg aacgggttgg
     5161 catggattgt aggcgccgcc ctataccttg tctgcctccc cgcgttgcgt cgcggtgcat
     5221 ggagccgggc cacctcgacc tgaatggaag ccggcggcac ctcgctaacg gattcaccac
     5281 tccaagaatt ggagccaatc aattcttgcg gagaactgtg aatgcgcaaa ccaacccttg
     5341 gcagaacata tccatcgcgt ccgccatctc cagcagccgc acgcggcgca tctcgggcag
     5401 cgttgggtcc tggccacggg tgcgcatgat cgtgctcctg tcgttgagga cccggctagg
     5461 ctggcggggt tgccttactg gttagcagaa tgaatcaccg atacgcgagc gaacgtgaag
     5521 cgactgctgc tgcaaaacgt ctgcgacctg agcaacaaca tgaatggtct tcggtttccg
     5581 tgtttcgtaa agtctggaaa cgcggaagtc agcgccctgc accattatgt tccggatctg
     5641 catcgcagga tgctgctggc taccctgtgg aacacctaca tctgtattaa cgaagcgctg
     5701 gcattgaccc tgagtgattt ttctctggtc ccgccgcatc cataccgcca gttgtttacc
     5761 ctcacaacgt tccagtaacc gggcatgttc atcatcagta acccgtatcg tgagcatcct
     5821 ctctcgtttc atcggtatca ttacccccat gaacagaaat cccccttaca cggaggcatc
     5881 agtgaccaaa caggaaaaaa ccgcccttaa catggcccgc tttatcagaa gccagacatt
     5941 aacgcttctg gagaaactca acgagctgga cgcggatgaa caggcagaca tctgtgaatc
     6001 gcttcacgac cacgctgatg agctttaccg cagctgcctc gcgcgtttcg gtgatgacgg
     6061 tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt aagcggatgc
     6121 cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggcgcagc
     6181 catgacccag tcacgtagcg atagcggagt gtatactggc ttaactatgc ggcatcagag
     6241 cagattgtac tgagagtgca ccattgcggt gtgaaatacc gcacagatgc gtaaggagaa
     6301 aataccgcat caggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     6361 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     6421 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     6481 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     6541 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     6601 ctggaagctc cctcgt
//
