LOCUS       pNQ059_S1-pE-SUM        7409 bp DNA     circular SYN 29-JUN-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7409)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jun 29, 2025 from SnapGene 7.2.0
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7409
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(865..1947)
                     /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"
     promoter        complement(1948..2025)
                     /gene="lacI"
                     /label=lacI promoter
     promoter        2334..2352
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2353..2377
                     /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             2392..2414
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             2427..2444
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             2457..2744
                     /codon_start=1
                     /gene="S. cerevisiae SMT3 (truncated and modified)"
                     /product="cleavable ubiquitin-like protein tag"
                     /label=SUMO
                     /note="modified to prevent cleavage by eukaryotic SUMO 
                     proteases"
                     /translation="DSEVNQEAKPEVKPEVKPETHINLKVSDGSSEIFFKIKKTTPLRR
                     LMEAFAKRQGKEMDSLRFLYDGIRIQADQAPEDLDMEDNDIIEAHREQIGG"
     CDS             4470..4487
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      4554..4601
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      4638..5093
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        5120..5224
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5225..5293,5294..6085)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      6256..6844
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    7030..7172
                     /label=bom
                     /note="basis of mobility region from pBR322"
ORIGIN
        1 gtctggcttc tgataaagcg ggccatgtta agggcggttt tttcctgttt ggtcactgat
       61 gcctccgtgt aagggggatt tctgttcatg ggggtaatga taccgatgaa acgagagagg
      121 atgctcacga tacgggttac tgatgatgaa catgcccggt tactggaacg ttgtgagggt
      181 aaacaactgg cggtatggat gcggcgggac cagagaaaaa tcactcaggg tcaatgccag
      241 cgcttcgtta atacagatgt aggtgttcca cagggtagcc agcagcatcc tgcgatgcag
      301 atccggaaca taatggtgca gggcgctgac ttccgcgttt ccagacttta cgaaacacgg
      361 aaaccgaaga ccattcatgt tgttgctcag gtcgcagacg ttttgcagca gcagtcgctt
      421 cacgttcgct cgcgtatcgg tgattcattc tgctaaccag taaggcaacc ccgccagcct
      481 agccgggtcc tcaacgacag gagcacgatc atgcgcaccc gtggggccgc catgccggcg
      541 ataatggcct gcttctcgcc gaaacgtttg gtggcgggac cagtgacgaa ggcttgagcg
      601 agggcgtgca agattccgaa taccgcaagc gacaggccga tcatcgtcgc gctccagcga
      661 aagcggtcct cgccgaaaat gacccagagc gctgccggca cctgtcctac gagttgcatg
      721 ataaagaaga cagtcataag tgcggcgacg atagtcatgc cccgcgccca ccggaaggag
      781 ctgactgggt tgaaggctct caagggcatc ggtcgagatc ccggtgccta atgagtgagc
      841 taacttacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
      901 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgccag
      961 ggtggttttt cttttcacca gtgagacggg caacagctga ttgcccttca ccgcctggcc
     1021 ctgagagagt tgcagcaagc ggtccacgct ggtttgcccc agcaggcgaa aatcctgttt
     1081 gatggtggtt aacggcggga tataacatga gctgtcttcg gtatcgtcgt atcccactac
     1141 cgagatatcc gcaccaacgc gcagcccgga ctcggtaatg gcgcgcattg cgcccagcgc
     1201 catctgatcg ttggcaacca gcatcgcagt gggaacgatg ccctcattca gcatttgcat
     1261 ggtttgttga aaaccggaca tggcactcca gtcgccttcc cgttccgcta tcggctgaat
     1321 ttgattgcga gtgagatatt tatgccagcc agccagacgc agacgcgccg agacagaact
     1381 taatgggccc gctaacagcg cgatttgctg gtgacccaat gcgaccagat gctccacgcc
     1441 cagtcgcgta ccgtcttcat gggagaaaat aatactgttg atgggtgtct ggtcagagac
     1501 atcaagaaat aacgccggaa cattagtgca ggcagcttcc acagcaatgg catcctggtc
     1561 atccagcgga tagttaatga tcagcccact gacgcgttgc gcgagaagat tgtgcaccgc
     1621 cgctttacag gcttcgacgc cgcttcgttc taccatcgac accaccacgc tggcacccag
     1681 ttgatcggcg cgagatttaa tcgccgcgac aatttgcgac ggcgcgtgca gggccagact
     1741 ggaggtggca acgccaatca gcaacgactg tttgcccgcc agttgttgtg ccacgcggtt
     1801 gggaatgtaa ttcagctccg ccatcgccgc ttccactttt tcccgcgttt tcgcagaaac
     1861 gtggctggcc tggttcacca cgcgggaaac ggtctgataa gagacaccgg catactctgc
     1921 gacatcgtat aacgttactg gtttcacatt caccaccctg aattgactct cttccgggcg
     1981 ctatcatgcc ataccgcgaa aggttttgcg ccattcgatg gtgtccggga tctcgacgct
     2041 ctcccttatg cgactcctgc attaggaagc agcccagtag taggttgagg ccgttgagca
     2101 ccgccgccgc aaggaatggt gcatgcaagg agatggcgcc caacagtccc ccggccacgg
     2161 ggcctgccac catacccacg ccgaaacaag cgctcatgag cccgaagtgg cgagcccgat
     2221 cttccccatc ggtgatgtcg gcgatatagg cgccagcaac cgcacctgtg gcgccggtga
     2281 tgccggccac gatgcgtccg gcgtagagga tcgagatctc gatcccgcga aattaatacg
     2341 actcactata ggggaattgt gagcggataa caattcccct ctagaaataa ttttgtttaa
     2401 ctttaagaag gagatatacc atgggtcatc accatcatca tcacgggtcc ctgcaggact
     2461 cagaagtcaa tcaagaagct aagccagagg tcaagccaga agtcaagcct gagactcaca
     2521 tcaatttaaa ggtgtccgat ggatcttcag agatcttctt caagatcaaa aagaccactc
     2581 ctttaagaag gctgatggaa gcgttcgcta aaagacaggg taaggaaatg gactccttaa
     2641 gattcttgta cgacggtatt agaattcaag ctgatcaggc ccctgaagat ttggacatgg
     2701 aggataacga tattattgag gctcaccgcg aacagattgg aggtatgact gaatcttttg
     2761 ctcaactctt tgaagagtcc ttaaaagaaa tcgaaacccg cccgggttct atcgttcgtg
     2821 gcgttgttgt tgctatcgac aaagacgtag tactggttga cgctggtctg aaatctgagt
     2881 ccgccatccc ggctgagcag ttcaaaaacg cccagggcga gctggaaatc caggtaggtg
     2941 acgaagttga cgttgctctg gacgcagtag aagacggctt cggtgaaact ctgctgtccc
     3001 gtgagaaagc taaacgtcac gaagcctgga tcacgctgga aaaagcttac gaagatgctg
     3061 aaactgttac cggtgttatc aacggcaaag ttaagggcgg cttcactgtt gagctgaacg
     3121 gtattcgtgc gttcctgcca ggttctctgg tagacgttcg tccggtgcgt gacactctgc
     3181 acctggaagg caaagagctt gaatttaaag taatcaagct ggatcagaag cgcaacaacg
     3241 ttgttgtttc tcgtcgtgcc gttatcgaat ccgaaaacag cgcagagcgc gatcagctgc
     3301 tggaaaacct gcaggaaggc atggaagtta aaggtatcgt taagaacctc actgactacg
     3361 gtgcattcgt tgatctgggc ggcgttgacg gcctgctgca catcactgac atggcctgga
     3421 aacgcgttaa gcatccgagc gaaatcgtca acgtgggcga cgaaatcact gttaaagtgc
     3481 tgaagttcga ccgcgaacgt acccgtgtat ccctgggcct gaaacagctg ggcgaagatc
     3541 cgtgggtagc tatcgctaaa cgttatccgg aaggtaccaa actgactggt cgcgtgacca
     3601 acctgaccga ctacggctgc ttcgttgaaa tcgaagaagg cgttgaaggc ctggtacacg
     3661 tttccgaaat ggactggacc aacaaaaaca tccacccgtc caaagttgtt aacgttggcg
     3721 atgtagtgga agttatggtt ctggatatcg acgaagaacg tcgtcgtatc tccctgggtc
     3781 tgaaacagtg caaagctaac ccgtggcagc agttcgcgga aacccacaac aagggcgacc
     3841 gtgttgaagg taaaatcaag tctatcactg acttcggtat cttcatcggc ttggacggcg
     3901 gcatcgacgg cctggttcac ctgtctgaca tctcctggaa cgttgcaggc gaagaagcag
     3961 ttcgtgaata caaaaaaggc gacgaaatcg ctgcagttgt tctgcaggtt gacgcagaac
     4021 gtgaacgtat ctccctgggc gttaaacagc tcgcagaaga tccgttcaac aactgggttg
     4081 ctctgaacaa gaaaggcgct atcgtaaccg gtaaagtaac tgcagttgac gctaaaggcg
     4141 caaccgtaga actggctgac ggcgttgaag gttacctgcg tgcttctgaa gcatcccgtg
     4201 accgcgttga agacgctacc ctggttctga gcgttggcga cgaagttgaa gctaaattca
     4261 ccggcgttga tcgtaaaaac cgcgcaatca gcctgtctgt tcgtgcgaaa gacgaagctg
     4321 acgagaaaga tgcaatcgca actgttaaca aacaggaaga tgcaaacttc tccaacaacg
     4381 caatggctga agctttcaaa gcagctaaag gcgagtaact agaggatccg aattcgagct
     4441 ccgtcgacaa gcttgcggcc gcactcgagc accaccacca ccaccactga gatccggctg
     4501 ctaacaaagc ccgaaaggaa gctgagttgg ctgctgccac cgctgagcaa taactagcat
     4561 aaccccttgg ggcctctaaa cgggtcttga ggggtttttt gctgaaagga ggaactatat
     4621 ccggattggc gaatgggacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     4681 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
     4741 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
     4801 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga
     4861 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     4921 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt
     4981 ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct
     5041 gatttaacaa aaatttaacg cgaattttaa caaaatatta acgtttacaa tttcaggtgg
     5101 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     5161 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     5221 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
     5281 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
     5341 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
     5401 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
     5461 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
     5521 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
     5581 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
     5641 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
     5701 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     5761 gatgcctgca gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
     5821 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
     5881 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     5941 ttctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     6001 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     6061 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
     6121 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     6181 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     6241 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     6301 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     6361 gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta
     6421 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     6481 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     6541 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     6601 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     6661 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     6721 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     6781 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     6841 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     6901 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
     6961 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
     7021 ggaagagcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat
     7081 atatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca gtatacactc
     7141 cgctatcgct acgtgactgg gtcatggctg cgccccgaca cccgccaaca cccgctgacg
     7201 cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg
     7261 ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgagg cagctgcggt
     7321 aaagctcatc agcgtggtcg tgaagcgatt cacagatgtc tgcctgttca tccgcgtcca
     7381 gctcgttgag tttctccaga agcgttaat
//
