LOCUS       pNQ055_pE-SUMO_M        7382 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 7382)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jun 29, 2025 from SnapGene 7.2.0
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7382
                     /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             4443..4460
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      4527..4574
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      4611..5066
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        5093..5197
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5198..5266,5267..6058)
                     /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      6229..6817
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    7003..7145
                     /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 caagtcgcga
     2761 agaaaattct ggtgacgtgc gcactgccgt acgctaacgg ctcaatccac ctcggccata
     2821 tgctggagca catccaggct gatgtctggg tccgttacca gcgaatgcgc ggccacgagg
     2881 tcaacttcat ctgcgccgac gatgcccacg gtacaccgat catgctgaaa gctcagcagc
     2941 ttggtatcac cccggagcag atgattggcg aaatgagtca ggagcatcag actgatttcg
     3001 caggctttaa catcagctat gacaactatc actcgacgca cagcgaagag aaccgccagt
     3061 tgtcagaact tatctactct cgcctgaaag aaaacggttt tattaaaaac cgcaccatct
     3121 ctcagctgta cgatccggaa aaaggcatgt tcctgccgga ccgttttgtg aaaggcacct
     3181 gcccgaaatg taaatccccg gatcaatacg gcgataactg cgaagtctgc ggcgcgacct
     3241 acagcccgac tgaactgatc gagccgaaat cggtggtttc tggcgctacg ccggtaatgc
     3301 gtgattctga acacttcttc tttgatctgc cctctttcag cgaaatgttg caggcatgga
     3361 cccgcagcgg tgcgttgcag gagcaggtgg caaataaaat gcaggagtgg tttgaatctg
     3421 gcctgcaaca gtgggatatc tcccgcgacg ccccttactt cggttttgaa attccgaacg
     3481 cgccgggcaa atatttctac gtctggctgg acgcaccgat tggctacatg ggttctttca
     3541 agaatctgtg cgacaagcgc ggcgacagcg taagcttcga tgaatactgg aagaaagact
     3601 ccaccgccga gctgtaccac ttcatcggta aagatattgt ttacttccac agcctgttct
     3661 ggcctgccat gctggaaggc agcaacttcc gcaagccgtc caacctgttt gttcatggct
     3721 atgtgacggt gaacggcgca aagatgtcca agtctcgcgg cacctttatt aaagccagca
     3781 cctggctgaa tcattttgac gcagacagcc tgcgttacta ctacactgcg aaactctctt
     3841 cgcgcattga tgatatcgat ctcaacctgg aagatttcgt tcagcgtgtg aatgccgata
     3901 tcgttaacaa agtggttaac ctggcctccc gtaatgcggg ctttatcaac aagcgttttg
     3961 acggcgtgct ggcaagcgaa ctggctgacc cgcagttgta caaaaccttc actgatgccg
     4021 ctgaagtgat tggtgaagcg tgggaaagcc gtgaatttgg taaagccgtg cgcgaaatca
     4081 tggcgctggc tgatctggct aaccgctatg tcgatgaaca ggctccgtgg gtggtggcga
     4141 aacaggaagg ccgcgatgcc gacctgcagg caatttgctc aatgggcatc aacctgttcc
     4201 gcgtgctgat gacttacctg aagccggtac tgccgaaact gaccgagcgt gcagaagcat
     4261 tcctcaatac ggaactgacc tgggatggta tccagcaacc gctgctgggc cacaaagtga
     4321 atccgttcaa ggcgctgtat aaccgcatcg atatgaggca ggttgaagca ctggtggaag
     4381 cctctaaata actagaggat ccgaattcga gctccgtcga caagcttgcg gccgcactcg
     4441 agcaccacca ccaccaccac tgagatccgg ctgctaacaa agcccgaaag gaagctgagt
     4501 tggctgctgc caccgctgag caataactag cataacccct tggggcctct aaacgggtct
     4561 tgaggggttt tttgctgaaa ggaggaacta tatccggatt ggcgaatggg acgcgccctg
     4621 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     4681 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     4741 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     4801 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg
     4861 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     4921 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt
     4981 gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt
     5041 taacaaaata ttaacgttta caatttcagg tggcactttt cggggaaatg tgcgcggaac
     5101 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     5161 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
     5221 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
     5281 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
     5341 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
     5401 cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca
     5461 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
     5521 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
     5581 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
     5641 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     5701 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gcagcaatgg caacaacgtt
     5761 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
     5821 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
     5881 tattgctgat aaatctggag ccggtgagcg tggttctcgc ggtatcattg cagcactggg
     5941 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
     6001 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
     6061 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     6121 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     6181 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     6241 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     6301 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     6361 gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     6421 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     6481 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     6541 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     6601 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     6661 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     6721 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     6781 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     6841 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
     6901 ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac
     6961 gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcctgatgc ggtattttct
     7021 ccttacgcat ctgtgcggta tttcacaccg catatatggt gcactctcag tacaatctgc
     7081 tctgatgccg catagttaag ccagtataca ctccgctatc gctacgtgac tgggtcatgg
     7141 ctgcgccccg acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg
     7201 catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac
     7261 cgtcatcacc gaaacgcgcg aggcagctgc ggtaaagctc atcagcgtgg tcgtgaagcg
     7321 attcacagat gtctgcctgt tcatccgcgt ccagctcgtt gagtttctcc agaagcgtta
     7381 at
//
