LOCUS       sequence-64173-e        4565 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4565)
  AUTHORS   Ponchon L, Catala M, Seijo B, El Khouri M, Dardel F, Nonin-Lecomte 
            S, Tisne C
  TITLE     Co-expression of RNA-protein complexes in Escherichia coli and 
            applications to RNA biology.
  JOURNAL   Nucleic Acids Res. 2013 Aug;41(15):e150. doi: 10.1093/nar/gkt576. 
            Epub 2013 Jun 26.
  PUBMED    23804766
REFERENCE   2  (bases 1 to 4565)
  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: "Nucleic 
            Acids Res."; date: "2013-08"; volume: "41(15):e150. doi"; pages: " 
            10.1093/nar/gkt576. Epub 2013 Jun 26"
COMMENT     Sequence Label: pProRNA
FEATURES             Location/Qualifiers
     source          1..4565
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        36..54
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     CDS             complement(87..1166)
                     /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     1128..1147
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(1167..1244)
                     /gene="lacI"
                     /label=lacI promoter
     promoter        1290..1308
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1309..1333
                     /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             1389..1406
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     promoter        join(1503..1508,1509..1524,1525..1531)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    1539..1555
                     /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)."
     promoter        complement(1792..1810)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1817..1834)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1817..1833)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1826..1848)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(2055..2074)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2454..2558
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2559..2627,2628..3419)
                     /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"
     primer_bind     complement(2777..2796)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3590..4178
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4079..4098
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4332..4349
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4466..4487
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4502..4507,4508..4525,4526..4532)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4540..4556
                     /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     join(4564..4565,1..15)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     join(4564..4565,1..15)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 ggaaacagct atgaccatga ttacgccaag ctcggaatta accctcacta aagggaacaa
       61 aagctgggta ccgggccccc cctcgactgc ccgctttcca gtcgggaaac ctgtcgtgcc
      121 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgccagg
      181 gtggtttttc ttttcaccag tgagacgggc aacagctgat tgcccttcac cgcctggccc
      241 tgagagagtt gcagcaagcg gtccacgctg gtttgcccca gcaggcgaaa atcctgtttg
      301 atggtggtta acggcgggat ataacatgag ctgtcttcgg tatcgtcgta tcccactacc
      361 gagatgtccg caccaacgcg cagcccggac tcggtaatgg cgcgcattgc gcccagcgcc
      421 atctgatcgt tggcaaccag catcgcagtg ggaacgatgc cctcattcag catttgcatg
      481 gtttgttgaa aaccggacat ggcactccag tcgccttccc gttccgctat cggctgaatt
      541 tgattgcgag tgagatattt atgccagcca gccagacgca gacgcgccga gacagaactt
      601 aatgggcccg ctaacagcgc gatttgctgg tgacccaatg cgaccagatg ctccacgccc
      661 agtcgcgtac cgtcttcatg ggagaaaata atactgttga tgggtgtctg gtcagagaca
      721 tcaagaaata acgccggaac attagtgcag gcagcttcca cagcaatggc atcctggtca
      781 tccagcggat agttaatgat cagcccactg acgcgttgcg cgagaagatt gtgcaccgcc
      841 gctttacagg cttcgacgcc gcttcgttct accatcgaca ccaccacgct ggcacccagt
      901 tgatcggcgc gagatttaat cgccgcgaca atttgcgacg gcgcgtgcag ggccagactg
      961 gaggtggcaa cgccaatcag caacgactgt ttgcccgcca gttgttgtgc cacgcggttg
     1021 ggaatgtaat tcagctccgc catcgccgct tccacttttt cccgcgtttt cgcagaaacg
     1081 tggctggcct ggttcaccac gcgggaaacg gtctgataag agacaccggc atactctgcg
     1141 acatcgtata acgttactgg tttcacattc accaccctga attgactctc ttccgggcgc
     1201 tatcatgcca taccgcgaaa ggttttgcgc cattcgatgg tgtccgggat ctcgacgctc
     1261 tcccttatgc gactcctgca ttaggaaatt aatacgactc actatagggg aattgtgagc
     1321 ggataacaat tcccctgtag aaataatttt gtttaacttt aataaggaga tataccatgg
     1381 gcagcagcca tcaccatcat caccacagcc aggatccgaa ttcgagctcg gcgcgcctgc
     1441 aggtcgacaa gcttgcggcc gcataatgct taagtcgaac agaaagtaat cgtattgtac
     1501 agttgacaat taatcatcgg ctcgtataat gtgtggaatt gtgagcggat aacaatttta
     1561 aggagatata catatggcag atctcaattg gatatcggcc ggccacgcga tcgctgacgt
     1621 cggtaccatc gaggtcgccc catcaaaaaa atattctcaa cataaaaaac tttgtgtaat
     1681 acttgtaacg ctgaattccc cgggctgcag atccttagcg aaagctaagg atttttttta
     1741 agcttgatgg gggcttagga tccggagctc caagctttag ctccaattcg ccctatagtg
     1801 agtcgtatta caattcactg gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg
     1861 ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt aatagcgaag
     1921 aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tggcctgtag
     1981 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     2041 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     2101 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     2161 cctcgacccc aaaaaacttg atttgggtga tggttcacgt agtgggccat cgccctgata
     2221 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     2281 aacttgaaca acactcaacc ctatctcggg ctattctttt gatttataag ggattttgcc
     2341 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa
     2401 caaaatatta acgtttacaa tttaaatcag gtggcacttt tcggggaaat gtgcgcggaa
     2461 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     2521 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg
     2581 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
     2641 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg
     2701 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga
     2761 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc
     2821 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag
     2881 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
     2941 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg
     3001 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
     3061 atgaagccat accaaacgac gagcgtgaca ccacgatgcc agcagcaatg gcaacaacgt
     3121 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact
     3181 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
     3241 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg
     3301 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
     3361 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac
     3421 tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat ttttaattta
     3481 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
     3541 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     3601 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     3661 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     3721 agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     3781 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     3841 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     3901 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     3961 tgagatacct acagcgtgag cattgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     4021 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     4081 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     4141 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     4201 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg
     4261 attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa
     4321 cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc
     4381 ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga
     4441 aagcgggcag tgagcgcaac gcaattaatg tgagttagct cactcattag gcaccccagg
     4501 ctttacactt tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc
     4561 acaca
//
