LOCUS       sequence-469254-        6643 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  over-express E. coli holA (Pol III delta subunit).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6643)
  AUTHORS   O'Donnell Lab
  TITLE     O'Donnell Lab Batch 1
  JOURNAL   unpublished
REFERENCE   2  (bases 1 to 6643)
  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            "unpublished"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 holA-pET(11a)(ACH-)
FEATURES             Location/Qualifiers
     source          1..6643
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(139..727)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(219..238)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(898..1689,1690..1758))
                     /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     1521..1540
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1759..1863)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1927..1945
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(2175..2222)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     2218..2236
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     protein_bind    3340..3364
                     /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(3364..3383)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3365..3383)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3467..3486)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        3692..3769
                     /gene="lacI"
                     /label=lacI promoter
     CDS             3770..4852
                     /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(3789..3808)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             6164..6355
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(6349..6371)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    6457..6596
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     6512..6531
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(6611..6628)
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
       61 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
      121 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
      181 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
      241 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
      301 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
      361 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
      421 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
      481 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
      541 gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc
      601 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
      661 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
      721 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
      781 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
      841 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
      901 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
      961 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     1021 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     1081 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     1141 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     1201 tgttgccatt gctgcaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     1261 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     1321 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     1381 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     1441 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     1501 ttgcccggcg tcaacacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     1561 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     1621 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     1681 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     1741 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     1801 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     1861 gcgcacattt ccccgaaaag tgccacctgc taagaaacca ttattatcat gacattaacc
     1921 tataaaaata ggcgtatcac gaggcccttt cgtcttcaag aattctcatg tttgacagct
     1981 tatcatcgac cagctttaat gcggtagttt atcacagtta aattgctaac gcagtcaggc
     2041 accgtgtatg aaatctaaca atgcgctcat cgtcatcctc ggcaccgtca ccctggatgc
     2101 tgtaggcata ggcttggtta tgccggtact gccgggcctc ttgcgggata tccggatata
     2161 gttcctcctt tcagcaaaaa acccctcaag acccgtttag aggccccaag gggttatgct
     2221 agttattgct cagcggtggc agcagccaac tcagcttcct ttcgggcttt gttagcagcc
     2281 ggatcctcaa ccatcgataa atacgtccgc caggggttta tggcacaaca gaagagataa
     2341 cccttccagc tctgcccaca ctgactgacc gtaatcttgt ttgagggtga gttccgttcg
     2401 tgtcaggagt tgcacggcct gacgtaactg cgtctgactt aagcgattta acgcctcgcc
     2461 catcatgccc cggcggttct gccatacccg atgcttatca aacaacgcac gcagtggcgt
     2521 atgggcagac tggcgtttca ggttaaccag taacaacagt tcacgttgta atgtgcgcaa
     2581 caaaataacc ggttcgctgc cttccagacg cagttgctga agaatatgca atgcgcgctt
     2641 actttttccc atcaacaaag catcaaccca atgaaaaggg gtgaaatgcg cggcatcatt
     2701 caccgcctgt tcaacgcgcg gtaatgtcaa tttgccgtct ggccagagca gcgataaacg
     2761 ctccagtgcc tgagccagcg ccagcaggtt accttcataa cagtagcaga gcacctgatt
     2821 tgccgcgtca tccagttcta agttgagctg ttttgcgcgc gcagcaaccc agcggggaag
     2881 ctgagcctgc tccggtgtct gacaggtcac ctgcacgctg cgattcgcaa gcgcagtaaa
     2941 ccaggcggca ttttcttgcg ctttgcttaa tttattaccg cggacgatca acagcaggtc
     3001 gtcatgcaga agtccggtga gtgtgagaag ttgctcattg atcgccgcat tcggtccgtt
     3061 ttctggtaac aacagcaata gcgtttgtcg actggcaaac agactcatag cctggcataa
     3121 cgaaaagatc gcattccagt cagtgttggg atcaatggaa aaagtgtggt gttcttcgaa
     3181 tccttgtgcc gcagctacct gacgaacagc gtcctggctt tcctgcaata acagaggatc
     3241 gttaccaagt aaaagatacg ccgcgcgcag cccttcattg agctgcgcgc ggagttgttc
     3301 cgggtacaac cgaatcatat ttatatctcc ttctagaggg gaattgttat ccgctcacaa
     3361 ttcccctata gtgagtcgta ttaatttcgc gggatcgaga tctcgatcct ctacgccgga
     3421 cgcatcgtgg ccggcatcac cggcgccaca ggtgcggttg ctggcgccta tatcgccgac
     3481 atcaccgatg gggaagatcg ggctcgccac ttcgggctca tgagcgcttg tttcggcgtg
     3541 ggtatggtgg caggccccgt ggccggggga ctgttgggcg ccatctcctt gcatgcacca
     3601 ttccttgcgg cggcggtgct caacggcctc aacctactac tgggctgctt cctaatgcag
     3661 gagtcgcata agggagagcg tcgagatccc ggacaccatc gaatggcgca aaacctttcg
     3721 cggtatggca tgatagcgcc cggaagagag tcaattcagg gtggtgaatg tgaaaccagt
     3781 aacgttatac gatgtcgcag agtatgccgg tgtctcttat cagaccgttt cccgcgtggt
     3841 gaaccaggcc agccacgttt ctgcgaaaac gcgggaaaaa gtggaagcgg cgatggcgga
     3901 gctgaattac attcccaacc gcgtggcaca acaactggcg ggcaaacagt cgttgctgat
     3961 tggcgttgcc acctccagtc tggccctgca cgcgccgtcg caaattgtcg cggcgattaa
     4021 atctcgcgcc gatcaactgg gtgccagcgt ggtggtgtcg atggtagaac gaagcggcgt
     4081 cgaagcctgt aaagcggcgg tgcacaatct tctcgcgcaa cgcgtcagtg ggctgatcat
     4141 taactatccg ctggatgacc aggatgccat tgctgtggaa gctgcctgca ctaatgttcc
     4201 ggcgttattt cttgatgtct ctgaccagac acccatcaac agtattattt tctcccatga
     4261 agacggtacg cgactgggcg tggagcatct ggtcgcattg ggtcaccagc aaatcgcgct
     4321 gttagcgggc ccattaagtt ctgtctcggc gcgtctgcgt ctggctggct ggcataaata
     4381 tctcactcgc aatcaaattc agccgatagc ggaacgggaa ggcgactgga gtgccatgtc
     4441 cggttttcaa caaaccatgc aaatgctgaa tgagggcatc gttcccactg cgatgctggt
     4501 tgccaacgat cagatggcgc tgggcgcaat gcgcgccatt accgagtccg ggctgcgcgt
     4561 tggtgcggat atctcggtag tgggatacga cgataccgaa gacagctcat gttatatccc
     4621 gccgttaacc accatcaaac aggattttcg cctgctgggg caaaccagcg tggaccgctt
     4681 gctgcaactc tctcagggcc aggcggtgaa gggcaatcag ctgttgcccg tctcactggt
     4741 gaaaagaaaa accaccctgg cgcccaatac gcaaaccgcc tctccccgcg cgttggccga
     4801 ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg
     4861 caattaatgt aagttagctc actcattagg caccgggatc tcgaccgatg cccttgagag
     4921 ccttcaaccc agtcagctcc ttccggtggg cgcggggcat gactatcgtc gccgcactta
     4981 tgactgtctt ctttatcatg caactcgtag gacaggtgcc ggcagcgctc tgggtcattt
     5041 tcggcgagga ccgctttcgc tggagcgcga cgatgatcgg cctgtcgctt gcggtattcg
     5101 gaatcttgca cgccctcgct caagccttcg tcactggtcc cgccaccaaa cgtttcggcg
     5161 agaagcaggc cattatcgcc ggcatggcgg ccgacgcgct gggctacgtc ttgctggcgt
     5221 tcgcgacgcg aggctggatg gccttcccca ttatgattct tctcgcttcc ggcggcatcg
     5281 ggatgcccgc gttgcaggcc atgctgtcca ggcaggtaga tgacgaccat cagggacagc
     5341 ttcaaggatc gctcgcggct cttaccagcc taacttcgat cattggaccg ctgatcgtca
     5401 cggcgattta tgccgcctcg gcgagcacat ggaacgggtt ggcatggatt gtaggcgccg
     5461 ccctatacct tgtctgcctc cccgcgttgc gtcgcggtgc atggagccgg gccacctcga
     5521 cctgaatgga agccggcggc acctcgctaa cggattcacc actccaagaa ttggagccaa
     5581 tcaattcttg cggagaactg tgaatgcgca aaccaaccct tggcagaaca tatccatcgc
     5641 gtccgccatc tccagcagcc gcacgcggcg catctcgggc agcgttgggt cctggccacg
     5701 ggtgcgcatg atcgtgctcc tgtcgttgag gacccggcta ggctggcggg gttgccttac
     5761 tggttagcag aatgaatcac cgatacgcga gcgaacgtga agcgactgct gctgcaaaac
     5821 gtctgcgacc tgagcaacaa catgaatggt cttcggtttc cgtgtttcgt aaagtctgga
     5881 aacgcggaag tcagcgccct gcaccattat gttccggatc tgcatcgcag gatgctgctg
     5941 gctaccctgt ggaacaccta catctgtatt aacgaagcgc tggcattgac cctgagtgat
     6001 ttttctctgg tcccgccgca tccataccgc cagttgttta ccctcacaac gttccagtaa
     6061 ccgggcatgt tcatcatcag taacccgtat cgtgagcatc ctctctcgtt tcatcggtat
     6121 cattaccccc atgaacagaa atccccctta cacggaggca tcagtgacca aacaggaaaa
     6181 aaccgccctt aacatggccc gctttatcag aagccagaca ttaacgcttc tggagaaact
     6241 caacgagctg gacgcggatg aacaggcaga catctgtgaa tcgcttcacg accacgctga
     6301 tgagctttac cgcagctgcc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat
     6361 gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg
     6421 tcagggcgcg tcagcgggtg ttggcgggtg tcggggcgca gccatgaccc agtcacgtag
     6481 cgatagcgga gtgtatactg gcttaactat gcggcatcag agcagattgt actgagagtg
     6541 caccattgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgct
     6601 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcg
//
