LOCUS       sequence-482112-        6413 bp    DNA     circular SYN 06-AUG-2025
DEFINITION  Over-express Ec dnaC subunit in E. coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6413)
  TITLE     O'Donnell lab plasmids 2025
REFERENCE   2  (bases 1 to 6413)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 6, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: Ec dnaC-pET(11a)
FEATURES             Location/Qualifiers
     source          1..6413
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(50..841,842..910))
                     /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     673..692
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(911..1015)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1083..1101
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(1128..1133,1134..1150,1151..1156)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     terminator      complement(1331..1378)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     1374..1392
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     RBS             2225..2247
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    2262..2286
                     /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(2286..2305)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2287..2305)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2389..2408)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        2614..2691
                     /gene="lacI"
                     /label=lacI promoter
     CDS             2692..3774
                     /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(2711..2730)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             5086..5277
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(5271..5293)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    5379..5518
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     5434..5453
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5533..5550)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5704..6292)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5784..5803)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
       61 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
      121 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
      181 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
      241 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
      301 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
      361 ttgctgcagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
      421 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
      481 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
      541 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
      601 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
      661 cgtcaacacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
      721 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
      781 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
      841 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
      901 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
      961 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     1021 ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata
     1081 aaaataggcg tatcacgagg ccctttcgtc ttcaagaatt ctcatgtttg acagcttatc
     1141 atcgataagc tttaatgcgg tagtttatca cagttaaatt gctaacgcag tcaggcaccg
     1201 tgtatgaaat ctaacaatgc gctcatcgtc atcctcggca ccgtcaccct ggatgctgta
     1261 ggcataggct tggttatgcc ggtactgccg ggcctcttgc gggatatccg gatatagttc
     1321 ctcctttcag caaaaaaccc ctcaagaccc gtttagaggc cccaaggggt tatgctagtt
     1381 attgctcagc ggtggcagca gccaactcag cttcctttcg ggctttgtta gcagccggat
     1441 ccaggtggaa ttgtgtcgca gtataccgcc agaaacgggg attaatactc tttacctgtt
     1501 acccggctac ggtagctatc ccagttgaag atcacccaca aactgttacc caggcgcata
     1561 cggtccatca cgcgttcgcc cagcaacttg gtcatctctt ccatattact gttggtcagc
     1621 atcccggttg ggcgtttgga agaagagcgg cgatcgacga tctggttgat gatcactttt
     1681 tcgtatttcg attcggtctg cacgccgatc tcatcgatca ccagcagatc aacgttgctc
     1741 agatcgttaa gcagttgttc ttcgctggta ccgctattcc tgaaggtatc tttcatcgcc
     1801 gacataatat cggccacggt gatgatcaat acggatttac cgcgtagcag cagctcgttg
     1861 cagattgccg ccgccagatg gtttttgccg gttcctggct taccagaaaa gatgaagctg
     1921 gcgatgttgc cgtcaaactc ttcgacatac tggcgcgctt tgcttaacgc attcatctgc
     1981 ccttcacact caacgcgata gttctcaaag gaacagttct gatgcagtgg acgaataccg
     2041 gagcggttaa aggtgcgctg cattttcatc gcccgatttt cacgttcgag agcggcggag
     2101 cggatcgctc cttgttcttt ttgccaggcc agtagttctt cacccgtttt gaatgcgggt
     2161 ttgatatggg caggcatcat tttttgcagg cgttgcatca ggtcgccaac gtttttcata
     2221 tatatctcct tcttaaagtt aaacaaaatt atttctagag gggaattgtt atccgctcac
     2281 aattccccta tagtgagtcg tattaatttc gcgggatcga gatctcgatc ctctacgccg
     2341 gacgcatcgt ggccggcatc accggcgcca caggtgcggt tgctggcgcc tatatcgccg
     2401 acatcaccga tggggaagat cgggctcgcc acttcgggct catgagcgct tgtttcggcg
     2461 tgggtatggt ggcaggcccc gtggccgggg gactgttggg cgccatctcc ttgcatgcac
     2521 cattccttgc ggcggcggtg ctcaacggcc tcaacctact actgggctgc ttcctaatgc
     2581 aggagtcgca taagggagag cgtcgagatc ccggacacca tcgaatggcg caaaaccttt
     2641 cgcggtatgg catgatagcg cccggaagag agtcaattca gggtggtgaa tgtgaaacca
     2701 gtaacgttat acgatgtcgc agagtatgcc ggtgtctctt atcagaccgt ttcccgcgtg
     2761 gtgaaccagg ccagccacgt ttctgcgaaa acgcgggaaa aagtggaagc ggcgatggcg
     2821 gagctgaatt acattcccaa ccgcgtggca caacaactgg cgggcaaaca gtcgttgctg
     2881 attggcgttg ccacctccag tctggccctg cacgcgccgt cgcaaattgt cgcggcgatt
     2941 aaatctcgcg ccgatcaact gggtgccagc gtggtggtgt cgatggtaga acgaagcggc
     3001 gtcgaagcct gtaaagcggc ggtgcacaat cttctcgcgc aacgcgtcag tgggctgatc
     3061 attaactatc cgctggatga ccaggatgcc attgctgtgg aagctgcctg cactaatgtt
     3121 ccggcgttat ttcttgatgt ctctgaccag acacccatca acagtattat tttctcccat
     3181 gaagacggta cgcgactggg cgtggagcat ctggtcgcat tgggtcacca gcaaatcgcg
     3241 ctgttagcgg gcccattaag ttctgtctcg gcgcgtctgc gtctggctgg ctggcataaa
     3301 tatctcactc gcaatcaaat tcagccgata gcggaacggg aaggcgactg gagtgccatg
     3361 tccggttttc aacaaaccat gcaaatgctg aatgagggca tcgttcccac tgcgatgctg
     3421 gttgccaacg atcagatggc gctgggcgca atgcgcgcca ttaccgagtc cgggctgcgc
     3481 gttggtgcgg atatctcggt agtgggatac gacgataccg aagacagctc atgttatatc
     3541 ccgccgttaa ccaccatcaa acaggatttt cgcctgctgg ggcaaaccag cgtggaccgc
     3601 ttgctgcaac tctctcaggg ccaggcggtg aagggcaatc agctgttgcc cgtctcactg
     3661 gtgaaaagaa aaaccaccct ggcgcccaat acgcaaaccg cctctccccg cgcgttggcc
     3721 gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa
     3781 cgcaattaat gtaagttagc tcactcatta ggcaccggga tctcgaccga tgcccttgag
     3841 agccttcaac ccagtcagct ccttccggtg ggcgcggggc atgactatcg tcgccgcact
     3901 tatgactgtc ttctttatca tgcaactcgt aggacaggtg ccggcagcgc tctgggtcat
     3961 tttcggcgag gaccgctttc gctggagcgc gacgatgatc ggcctgtcgc ttgcggtatt
     4021 cggaatcttg cacgccctcg ctcaagcctt cgtcactggt cccgccacca aacgtttcgg
     4081 cgagaagcag gccattatcg ccggcatggc ggccgacgcg ctgggctacg tcttgctggc
     4141 gttcgcgacg cgaggctgga tggccttccc cattatgatt cttctcgctt ccggcggcat
     4201 cgggatgccc gcgttgcagg ccatgctgtc caggcaggta gatgacgacc atcagggaca
     4261 gcttcaagga tcgctcgcgg ctcttaccag cctaacttcg atcattggac cgctgatcgt
     4321 cacggcgatt tatgccgcct cggcgagcac atggaacggg ttggcatgga ttgtaggcgc
     4381 cgccctatac cttgtctgcc tccccgcgtt gcgtcgcggt gcatggagcc gggccacctc
     4441 gacctgaatg gaagccggcg gcacctcgct aacggattca ccactccaag aattggagcc
     4501 aatcaattct tgcggagaac tgtgaatgcg caaaccaacc cttggcagaa catatccatc
     4561 gcgtccgcca tctccagcag ccgcacgcgg cgcatctcgg gcagcgttgg gtcctggcca
     4621 cgggtgcgca tgatcgtgct cctgtcgttg aggacccggc taggctggcg gggttgcctt
     4681 actggttagc agaatgaatc accgatacgc gagcgaacgt gaagcgactg ctgctgcaaa
     4741 acgtctgcga cctgagcaac aacatgaatg gtcttcggtt tccgtgtttc gtaaagtctg
     4801 gaaacgcgga agtcagcgcc ctgcaccatt atgttccgga tctgcatcgc aggatgctgc
     4861 tggctaccct gtggaacacc tacatctgta ttaacgaagc gctggcattg accctgagtg
     4921 atttttctct ggtcccgccg catccatacc gccagttgtt taccctcaca acgttccagt
     4981 aaccgggcat gttcatcatc agtaacccgt atcgtgagca tcctctctcg tttcatcggt
     5041 atcattaccc ccatgaacag aaatccccct tacacggagg catcagtgac caaacaggaa
     5101 aaaaccgccc ttaacatggc ccgctttatc agaagccaga cattaacgct tctggagaaa
     5161 ctcaacgagc tggacgcgga tgaacaggca gacatctgtg aatcgcttca cgaccacgct
     5221 gatgagcttt accgcagctg cctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac
     5281 atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc
     5341 cgtcagggcg cgtcagcggg tgttggcggg tgtcggggcg cagccatgac ccagtcacgt
     5401 agcgatagcg gagtgtatac tggcttaact atgcggcatc agagcagatt gtactgagag
     5461 tgcaccattg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg
     5521 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     5581 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     5641 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     5701 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     5761 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     5821 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     5881 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     5941 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     6001 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     6061 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     6121 gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt
     6181 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     6241 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     6301 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     6361 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aat
//
