LOCUS       sequence-482109-        7057 bp    DNA     circular SYN 06-AUG-2025
DEFINITION  Over-express Ec dnaB subunit in E. coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7057)
  TITLE     O'Donnell lab plasmids 2025
REFERENCE   2  (bases 1 to 7057)
  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 dnaB-pET
FEATURES             Location/Qualifiers
     source          1..7057
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(669..1460,1461..1529))
                     /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     1292..1311
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1530..1634)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1702..1720
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(1747..1752,1753..1769,1770..1775)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     terminator      complement(1950..1997)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     1993..2011
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     RBS             3488..3510
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    3525..3549
                     /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(3549..3568)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3550..3568)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3652..3671)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        3877..3954
                     /gene="lacI"
                     /label=lacI promoter
     CDS             3955..5037
                     /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(3974..3993)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             6349..6540
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(6534..6556)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    6642..6781
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     6697..6716
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(6796..6813)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(6967..7057,1..498))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(join(7047..7057,1..9))
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
       61 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
      121 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
      181 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
      241 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
      301 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt
      361 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
      421 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
      481 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
      541 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
      601 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
      661 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
      721 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
      781 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
      841 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
      901 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
      961 ttgcgcaacg ttgttgccat tgctgcaggc atcgtggtgt cacgctcgtc gtttggtatg
     1021 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     1081 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     1141 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     1201 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     1261 ccgagttgct cttgcccggc gtcaacacgg gataataccg cgccacatag cagaacttta
     1321 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     1381 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     1441 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     1501 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     1561 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     1621 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga aaccattatt
     1681 atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtct tcaagaattc
     1741 tcatgtttga cagcttatca tcgataagct ttaatgcggt agtttatcac agttaaattg
     1801 ctaacgcagt caggcaccgt gtatgaaatc taacaatgcg ctcatcgtca tcctcggcac
     1861 cgtcaccctg gatgctgtag gcataggctt ggttatgccg gtactgccgg gcctcttgcg
     1921 ggatatccgg atatagttcc tcctttcagc aaaaaacccc tcaagacccg tttagaggcc
     1981 ccaaggggtt atgctagtta ttgctcagcg gtggcagcag ccaactcagc ttcctttcgg
     2041 gctttgttag cagccggatc cttattattc gtcgtcgtac tgcggccccg catagttgtc
     2101 gaagcgcgac cattgaccgt taaaggtcag gcgtaccgtc ccgattgggc cgttacgttg
     2161 tttaccgata ataatttccg cgatgccttt taaatcactg ttttcgtgat acacctcatc
     2221 acgatagata aacatgatca agtccgcatc ctgctcgata gagccagatt cacgcaggtc
     2281 ggagttgacc gggcgtttgt cggcacgttg ttccagagaa cggttcaact gggacagcgc
     2341 caccaccggc acgttcagtt ctttcgccag tgctttcagc gagcgagaga tttctgcaat
     2401 ttccagcgta cggttatcgg aaagcgccgg tacgcgcatc agttgcaggt agtcgatcat
     2461 gataagcccg atgccgccgt gttcacgggc aatacggcgt gcgcgggaac gcacttccgt
     2521 tggcgtcagg ccggaggagt catcgatata gatattgcgt ttttcgagca aaatacccat
     2581 ggtgccggaa atgcgcgccc agtcttcgtc atcgagctgc ccggtacgga ttttagtctg
     2641 gtcaacgcgc gacagcgacg ccagagaacg catcatgatc tgttctgatg gcatctccag
     2701 cgagaagata agtaccggtt tatcctgcaa catcgccgcg ttttcgacga ggttcatcgc
     2761 aaatgttgtt ttacccatcg acggacgcgc ggcgacgatg atcaaatccg acggctgcaa
     2821 gccagcggtt tttttgttga gatcgtcata accggtgttt accccggtaa cgccatcgtg
     2881 tggctgctga aacaactgct caatacgcgc cacggttgcg tcgagcacat cggcgatgtt
     2941 cttcggccct tcgtctttgt tcgcacgact ttcggcaatt ttaaagacgc gggattcagc
     3001 cagatccagc agatcttcgc tggtacgccc ctgcggatca aaaccagctt cggcaatctc
     3061 attcgcaacc gagatcatct cacggacaac ggcacgttca cgcacgatgt ccgcataggc
     3121 actgatgtta gccgcacttg gcgtattttt tgacagctct gccagataag caaaaccacc
     3181 gacgctatcg agttgcccct ggcgttccag cgattccgca agagtaatca gatcgatagg
     3241 gctaccgctt tcctgcaaac gcgccatttc agtaaagata tgacggtgtg ggcgggtgta
     3301 aaaatcgtct gctaccacac gctcggctac atcatcccag cgttcgttat ctagcattaa
     3361 accgcccaac accgactgct ccgcttcgat cgagtgcgga ggcactttca gcccggcaac
     3421 ttgtggatcg cgttcgcggg gttcagcctg ctgtttgttg aagggtttat ttcctgccat
     3481 atgtatatct ccttcttaaa gttaaacaaa attatttcta gaggggaatt gttatccgct
     3541 cacaattccc ctatagtgag tcgtattaat ttcgcgggat cgagatctcg atcctctacg
     3601 ccggacgcat cgtggccggc atcaccggcg ccacaggtgc ggttgctggc gcctatatcg
     3661 ccgacatcac cgatggggaa gatcgggctc gccacttcgg gctcatgagc gcttgtttcg
     3721 gcgtgggtat ggtggcaggc cccgtggccg ggggactgtt gggcgccatc tccttgcatg
     3781 caccattcct tgcggcggcg gtgctcaacg gcctcaacct actactgggc tgcttcctaa
     3841 tgcaggagtc gcataaggga gagcgtcgag atcccggaca ccatcgaatg gcgcaaaacc
     3901 tttcgcggta tggcatgata gcgcccggaa gagagtcaat tcagggtggt gaatgtgaaa
     3961 ccagtaacgt tatacgatgt cgcagagtat gccggtgtct cttatcagac cgtttcccgc
     4021 gtggtgaacc aggccagcca cgtttctgcg aaaacgcggg aaaaagtgga agcggcgatg
     4081 gcggagctga attacattcc caaccgcgtg gcacaacaac tggcgggcaa acagtcgttg
     4141 ctgattggcg ttgccacctc cagtctggcc ctgcacgcgc cgtcgcaaat tgtcgcggcg
     4201 attaaatctc gcgccgatca actgggtgcc agcgtggtgg tgtcgatggt agaacgaagc
     4261 ggcgtcgaag cctgtaaagc ggcggtgcac aatcttctcg cgcaacgcgt cagtgggctg
     4321 atcattaact atccgctgga tgaccaggat gccattgctg tggaagctgc ctgcactaat
     4381 gttccggcgt tatttcttga tgtctctgac cagacaccca tcaacagtat tattttctcc
     4441 catgaagacg gtacgcgact gggcgtggag catctggtcg cattgggtca ccagcaaatc
     4501 gcgctgttag cgggcccatt aagttctgtc tcggcgcgtc tgcgtctggc tggctggcat
     4561 aaatatctca ctcgcaatca aattcagccg atagcggaac gggaaggcga ctggagtgcc
     4621 atgtccggtt ttcaacaaac catgcaaatg ctgaatgagg gcatcgttcc cactgcgatg
     4681 ctggttgcca acgatcagat ggcgctgggc gcaatgcgcg ccattaccga gtccgggctg
     4741 cgcgttggtg cggatatctc ggtagtggga tacgacgata ccgaagacag ctcatgttat
     4801 atcccgccgt taaccaccat caaacaggat tttcgcctgc tggggcaaac cagcgtggac
     4861 cgcttgctgc aactctctca gggccaggcg gtgaagggca atcagctgtt gcccgtctca
     4921 ctggtgaaaa gaaaaaccac cctggcgccc aatacgcaaa ccgcctctcc ccgcgcgttg
     4981 gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg
     5041 caacgcaatt aatgtaagtt agctcactca ttaggcaccg ggatctcgac cgatgccctt
     5101 gagagccttc aacccagtca gctccttccg gtgggcgcgg ggcatgacta tcgtcgccgc
     5161 acttatgact gtcttcttta tcatgcaact cgtaggacag gtgccggcag cgctctgggt
     5221 cattttcggc gaggaccgct ttcgctggag cgcgacgatg atcggcctgt cgcttgcggt
     5281 attcggaatc ttgcacgccc tcgctcaagc cttcgtcact ggtcccgcca ccaaacgttt
     5341 cggcgagaag caggccatta tcgccggcat ggcggccgac gcgctgggct acgtcttgct
     5401 ggcgttcgcg acgcgaggct ggatggcctt ccccattatg attcttctcg cttccggcgg
     5461 catcgggatg cccgcgttgc aggccatgct gtccaggcag gtagatgacg accatcaggg
     5521 acagcttcaa ggatcgctcg cggctcttac cagcctaact tcgatcattg gaccgctgat
     5581 cgtcacggcg atttatgccg cctcggcgag cacatggaac gggttggcat ggattgtagg
     5641 cgccgcccta taccttgtct gcctccccgc gttgcgtcgc ggtgcatgga gccgggccac
     5701 ctcgacctga atggaagccg gcggcacctc gctaacggat tcaccactcc aagaattgga
     5761 gccaatcaat tcttgcggag aactgtgaat gcgcaaacca acccttggca gaacatatcc
     5821 atcgcgtccg ccatctccag cagccgcacg cggcgcatct cgggcagcgt tgggtcctgg
     5881 ccacgggtgc gcatgatcgt gctcctgtcg ttgaggaccc ggctaggctg gcggggttgc
     5941 cttactggtt agcagaatga atcaccgata cgcgagcgaa cgtgaagcga ctgctgctgc
     6001 aaaacgtctg cgacctgagc aacaacatga atggtcttcg gtttccgtgt ttcgtaaagt
     6061 ctggaaacgc ggaagtcagc gccctgcacc attatgttcc ggatctgcat cgcaggatgc
     6121 tgctggctac cctgtggaac acctacatct gtattaacga agcgctggca ttgaccctga
     6181 gtgatttttc tctggtcccg ccgcatccat accgccagtt gtttaccctc acaacgttcc
     6241 agtaaccggg catgttcatc atcagtaacc cgtatcgtga gcatcctctc tcgtttcatc
     6301 ggtatcatta cccccatgaa cagaaatccc ccttacacgg aggcatcagt gaccaaacag
     6361 gaaaaaaccg cccttaacat ggcccgcttt atcagaagcc agacattaac gcttctggag
     6421 aaactcaacg agctggacgc ggatgaacag gcagacatct gtgaatcgct tcacgaccac
     6481 gctgatgagc tttaccgcag ctgcctcgcg cgtttcggtg atgacggtga aaacctctga
     6541 cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa
     6601 gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gcgcagccat gacccagtca
     6661 cgtagcgata gcggagtgta tactggctta actatgcggc atcagagcag attgtactga
     6721 gagtgcacca ttgcggtgtg aaataccgca cagatgcgta aggagaaaat accgcatcag
     6781 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     6841 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     6901 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     6961 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     7021 gaggtggcga aacccgacag gactataaag ataccag
//
