LOCUS       Exported                7272 bp DNA     circular SYN 16-SEP-2025
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7272)
  AUTHORS   11111111
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 7272)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Sep 16, 2025 from SnapGene 6.0.2
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..7272
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     source          join(2535..7272,1..2534)
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        130..148
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    149..173
                     /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)."
     CDS             218..1702
                     /codon_start=1
                     /gene="nusA"
                     /product="transcription elongation protein (N utilization
                     substance protein A)"
                     /label=NusA
                     /note="highly soluble in E. coli"
                     /translation="MNKEILAVVEAVSNEKALPREKIFEALESALATATKKKYEQEIDV
                     RVQIDRKSGDFDTFRRWLVVDEVTQPTKEITLEAARYEDESLNLGDYVEDQIESVTFDR
                     ITTQTAKQVIVQKVREAERAMVVDQFREHEGEIITGVVKKVNRDNISLDLGNNAEAVIL
                     REDMLPRENFRPGDRVRGVLYSVRPEARGAQLFVTRSKPEMLIELFRIEVPEIGEEVIE
                     IKAAARDPGSRAKIAVKTNDKRIDPVGACVGMRGARVQAVSTELGGERIDIVLWDDNPA
                     QFVINAMAPADVASIVVDEDKHTMDIAVEAGNLAQAIGRNGQNVRLASQLSGWELNVMT
                     VDDLQAKHQAEAHAAIDTFTKYLDIDEDFATVLVEEGFSTLEELAYVPMKELLEIEGLD
                     EPTVEALRERAKNALATIAQAQEESLGDNKPADDLLNLEGVDRDLAFKLAARGVCTLED
                     LAEQGIDDLADIEGLTDEKAGALIMAARNICWFGDEA"
     CDS             1718..1735
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1745..1789
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     CDS             1805..1822
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             1841..1855
                     /codon_start=1
                     /product="enterokinase recognition and cleavage site"
                     /label=enterokinase site
                     /translation="DDDDK"
     CDS             1968..2000
                     /codon_start=1
                     /product="HSV (herpes simplex virus) epitope tag"
                     /label=HSV tag
                     /translation="QPELAPEDPED"
     CDS             2007..2024
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      2190..2276
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      2368..2395
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     terminator      2463..2510
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2547..3002
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     CDS             complement(3117..3977)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(3978..4070)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4151..4739
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    4925..5064
                     /label=bom
                     /note="basis of mobility region from pBR322"
     CDS             complement(5166..5357)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5929..7011)
                     /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        7012..7089
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 ggcgagcccg atcttcccca tcggtgatgt cggcgatata ggcgccagca accgcacctg
       61 tggcgccggt gatgccggcc acgatgcgtc cggcgtagag gatcgagatc gatctcgatc
      121 ccgcgaaatt aatacgactc actatagggg aattgtgagc ggataacaat tcccctctag
      181 aaataatttt gtttaacttt aagaaggaga tatacatatg aacaaagaaa ttttggctgt
      241 agttgaagcc gtatccaatg aaaaggcgct acctcgcgag aagattttcg aagcattgga
      301 aagcgcgctg gcgacagcaa caaagaaaaa atatgaacaa gagatcgacg tccgcgtaca
      361 gatcgatcgc aaaagcggtg attttgacac tttccgtcgc tggttagttg ttgatgaagt
      421 cacccagccg accaaggaaa tcacccttga agccgcacgt tatgaagatg aaagcctgaa
      481 cctgggcgat tacgttgaag atcagattga gtctgttacc tttgaccgta tcactaccca
      541 gacggcaaaa caggttatcg tgcagaaagt gcgtgaagcc gaacgtgcga tggtggttga
      601 tcagttccgt gaacacgaag gtgaaatcat caccggcgtg gtgaaaaaag taaaccgcga
      661 caacatctct ctggatctgg gcaacaacgc tgaagccgtg atcctgcgcg aagatatgct
      721 gccgcgtgaa aacttccgcc ctggcgaccg cgttcgtggc gtgctctatt ccgttcgccc
      781 ggaagcgcgt ggcgcgcaac tgttcgtcac tcgttccaag ccggaaatgc tgatcgaact
      841 gttccgtatt gaagtgccag aaatcggcga agaagtgatt gaaattaaag cagcggctcg
      901 cgatccgggt tctcgtgcga aaatcgcggt gaaaaccaac gataaacgta tcgatccggt
      961 aggtgcttgc gtaggtatgc gtggcgcgcg tgttcaggcg gtgtctactg aactgggtgg
     1021 cgagcgtatc gatatcgtcc tgtgggatga taacccggcg cagttcgtga ttaacgcaat
     1081 ggcaccggca gacgttgctt ctatcgtggt ggatgaagat aaacacacca tggacatcgc
     1141 cgttgaagcc ggtaatctgg cgcaggcgat tggccgtaac ggtcagaacg tgcgtctggc
     1201 ttcgcaactg agcggttggg aactcaacgt gatgaccgtt gacgacctgc aagctaagca
     1261 tcaggcggaa gcgcacgcag cgatcgacac cttcaccaaa tatctcgaca tcgacgaaga
     1321 cttcgcgact gttctggtag aagaaggctt ctcgacgctg gaagaattgg cctatgtgcc
     1381 gatgaaagag ctgttggaaa tcgaaggcct tgatgagccg accgttgaag cactgcgcga
     1441 gcgtgctaaa aatgcactgg ccaccattgc acaggcccag gaagaaagcc tcggtgataa
     1501 caaaccggct gacgatctgc tgaaccttga aggggtagat cgtgatttgg cattcaaact
     1561 ggccgcccgt ggcgtttgta cgctggaaga tctcgccgaa cagggcattg atgatctggc
     1621 tgatatcgaa gggttgaccg acgaaaaagc cggagcactg attatggctg cccgtaatat
     1681 ttgctggttc ggtgacgaag cgactagtgg ttctggtcat caccatcacc atcactccgc
     1741 gggtaaagaa accgctgctg cgaaatttga acgccagcac atggactcgc caccgccaac
     1801 tggtctggtc ccccggggca gcgcgggttc tggtacgatt gatgacgacg acaagagtcc
     1861 gggagctcgt ggatccgaat tctgtacagg cgcgcctgca ggacgtcgac ggtaccatcg
     1921 atacgcgttc gaagcttgcg gccgcacagc tgtatacacg tgcaagccag ccagaactcg
     1981 ctcctgaaga cccagaggat ctcgagcacc accaccacca ccactaatgt taattaagtt
     2041 gggcgttcct aggctgataa aacagaattt gcctggcggc agtagcgcgg tggtcccacc
     2101 tgaccccatg ccgaactcag aagtgaaacg ccgtagcgcc gatggtagtg tggggtctcc
     2161 ccatgcgaga gtagggaact gccaggcatc aaataaaacg aaaggctcag tcgaaagact
     2221 gggcctttcg ttttatctgt tgtttgtcgg tgaacgctct cctgagtagg acaaatccgc
     2281 cgggagcgga tttgaacgtt gcgaagcaac ggcccggagg gtggcgggca ggacgcccgc
     2341 cataaactgc caggcatcaa attaagcaga aggccatcct gacggatggc ctttttgcgt
     2401 ttctacaaac tcttttgttt atttttctaa atacattcaa atatgtatcc gctgagcaat
     2461 aactagcata accccttggg gcctctaaac gggtcttgag gggttttttg ctgaaaggag
     2521 gaactatatc cggattggcg aatgggacgc gccctgtagc ggcgcattaa gcgcggcggg
     2581 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt
     2641 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     2701 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     2761 ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
     2821 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc
     2881 tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct attggttaaa
     2941 aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa cgtttacaat
     3001 ttctggcggc acgatggcat gagattatca aaaaggatct tcacctagat ccttttaaat
     3061 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     3121 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     3181 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     3241 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     3301 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     3361 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     3421 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     3481 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     3541 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     3601 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     3661 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     3721 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     3781 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     3841 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     3901 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     3961 aaatgttgaa tactcatact cttccttttt caatcatgat tgaagcattt atcagggtta
     4021 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggtcatga
     4081 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     4141 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     4201 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     4261 taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag
     4321 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     4381 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     4441 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     4501 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     4561 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     4621 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     4681 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     4741 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
     4801 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg
     4861 ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag
     4921 agcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcaatggtg
     4981 cactctcagt acaatctgct ctgatgccgc atagttaagc cagtatacac tccgctatcg
     5041 ctacgtgact gggtcatggc tgcgccccga cacccgccaa cacccgctga cgcgccctga
     5101 cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc
     5161 atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga ggcagctgcg gtaaagctca
     5221 tcagcgtggt cgtgaagcga ttcacagatg tctgcctgtt catccgcgtc cagctcgttg
     5281 agtttctcca gaagcgttaa tgtctggctt ctgataaagc gggccatgtt aagggcggtt
     5341 ttttcctgtt tggtcactga tgcctccgtg taagggggat ttctgttcat gggggtaatg
     5401 ataccgatga aacgagagag gatgctcacg atacgggtta ctgatgatga acatgcccgg
     5461 ttactggaac gttgtgaggg taaacaactg gcggtatgga tgcggcggga ccagagaaaa
     5521 atcactcagg gtcaatgcca gcgcttcgtt aatacagatg taggtgttcc acagggtagc
     5581 cagcagcatc ctgcgatgca gatccggaac ataatggtgc agggcgctga cttccgcgtt
     5641 tccagacttt acgaaacacg gaaaccgaag accattcatg ttgttgctca ggtcgcagac
     5701 gttttgcagc agcagtcgct tcacgttcgc tcgcgtatcg gtgattcatt ctgctaacca
     5761 gtaaggcaac cccgccagcc tagccgggtc ctcaacgaca ggagcacgat catgctagtc
     5821 atgccccgcg cccaccggaa ggagctgact gggttgaagg ctctcaaggg catcggtcga
     5881 gatcccggtg cctaatgagt gagctaactt acattaattg cgttgcgctc actgcccgct
     5941 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     6001 ggcggtttgc gtattgggcg ccagggtggt ttttcttttc accagtgaga cgggcaacag
     6061 ctgattgccc ttcaccgcct ggccctgaga gagttgcagc aagcggtcca cgctggtttg
     6121 ccccagcagg cgaaaatcct gtttgatggt ggttaacggc gggatataac atgagctgtc
     6181 ttcggtatcg tcgtatccca ctaccgagat gtccgcacca acgcgcagcc cggactcggt
     6241 aatggcgcgc attgcgccca gcgccatctg atcgttggca accagcatcg cagtgggaac
     6301 gatgccctca ttcagcattt gcatggtttg ttgaaaaccg gacatggcac tccagtcgcc
     6361 ttcccgttcc gctatcggct gaatttgatt gcgagtgaga tatttatgcc agccagccag
     6421 acgcagacgc gccgagacag aacttaatgg gcccgctaac agcgcgattt gctggtgacc
     6481 caatgcgacc agatgctcca cgcccagtcg cgtaccgtct tcatgggaga aaataatact
     6541 gttgatgggt gtctggtcag agacatcaag aaataacgcc ggaacattag tgcaggcagc
     6601 ttccacagca atggcatcct ggtcatccag cggatagtta atgatcagcc cactgacgcg
     6661 ttgcgcgaga agattgtgca ccgccgcttt acaggcttcg acgccgcttc gttctaccat
     6721 cgacaccacc acgctggcac ccagttgatc ggcgcgagat ttaatcgccg cgacaatttg
     6781 cgacggcgcg tgcagggcca gactggaggt ggcaacgcca atcagcaacg actgtttgcc
     6841 cgccagttgt tgtgccacgc ggttgggaat gtaattcagc tccgccatcg ccgcttccac
     6901 tttttcccgc gttttcgcag aaacgtggct ggcctggttc accacgcggg aaacggtctg
     6961 ataagagaca ccggcatact ctgcgacatc gtataacgtt actggtttca cattcaccac
     7021 cctgaattga ctctcttccg ggcgctatca tgccataccg cgaaaggttt tgcgccattc
     7081 gatggtgtcc gggatctcga cgctctccct tatgcgactc ctgcattagg aagcagccca
     7141 gtagtaggtt gaggccgttg agcaccgccg ccgcaaggaa tggtgcatgc aaggagatgg
     7201 cgcccaacag tcccccggcc acggggcctg ccaccatacc cacgccgaaa caagcgctca
     7261 tgagcccgaa gt
//
