LOCUS       sequence-64176-e        3975 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 3975)
  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 3975)
  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: pACYCT2
FEATURES             Location/Qualifiers
     source          1..3975
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    3..27
                     /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             83..100
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     promoter        join(197..202,203..218,219..225)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    233..249
                     /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             334..378
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(416..434)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      430..477
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(697..1356)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     1271..1290
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(1357..1459)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(1851..1868)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1985..2529)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     complement(2065..2084)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2754..3836)
                     /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     3798..3817
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(3837..3914)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 ggggaattgt gagcggataa caattcccct gtagaaataa ttttgtttaa ctttaataag
       61 gagatatacc atgggcagca gccatcacca tcatcaccac agccaggatc cgaattcgag
      121 ctcggcgcgc ctgcaggtcg acaagcttgc ggccgcataa tgcttaagtc gaacagaaag
      181 taatcgtatt gtacagttga caattaatca tcggctcgta taatgtgtgg aattgtgagc
      241 ggataacaat tttaaggaga tatacatatg gcagatctca attggatatc ggccggccac
      301 gcgatcgctg acgtcggtac cctcgagtct ggtaaagaaa ccgctgctgc gaaatttgaa
      361 cgccagcaca tggactcgtc tactagcgca gcttaattaa cctaggctgc tgccaccgct
      421 gagcaataac tagcataacc ccttggggcc tctaaacggg tcttgagggg ttttttgctg
      481 aaacctcagg catttgagaa gcacacggtc acactgcttc cggtagtcaa taaaccggta
      541 aaccagcaat agacataagc ggctatttaa cgaccctgcc ctgaaccgac gaccgggtcg
      601 aatttgcttt cgaatttctg ccattcatcc gcttattatc acttattcag gcgtagcacc
      661 aggcgtttaa gggcaccaat aactgcctta aaaaaattac gccccgccct gccactcatc
      721 gcagtactgt tgtaattcat taagcattct gccgacatgg aagccatcac agacggcatg
      781 atgaacctga atcgccagcg gcatcagcac cttgtcgcct tgcgtataat atttgcccat
      841 agtgaaaacg ggggcgaaga agttgtccat attggccacg tttaaatcaa aactggtgaa
      901 actcacccag ggattggctg agacgaaaaa catattctca ataaaccctt tagggaaata
      961 ggccaggttt tcaccgtaac acgccacatc ttgcgaatat atgtgtagaa actgccggaa
     1021 atcgtcgtgg tattcactcc agagcgatga aaacgtttca gtttgctcat ggaaaacggt
     1081 gtaacaaggg tgaacactat cccatatcac cagctcaccg tctttcattg ccatacggaa
     1141 ctccggatga gcattcatca ggcgggcaag aatgtgaata aaggccggat aaaacttgtg
     1201 cttatttttc tttacggtct ttaaaaaggc cgtaatatcc agctgaacgg tctggttata
     1261 ggtacattga gcaactgact gaaatgcctc aaaatgttct ttacgatgcc attgggatat
     1321 atcaacggtg gtatatccag tgattttttt ctccatttta gcttccttag ctcctgaaaa
     1381 tctcgataac tcaaaaaata cgcccggtag tgatcttatt tcattatggt gaaagttgga
     1441 acctcttacg tgccgatcaa cgtctcattt tcgccaaaag ttggcccagg gcttcccggt
     1501 atcaacaggg acaccaggat ttatttattc tgcgaagtga tcttccgtca caggtattta
     1561 ttcggcgcaa agtgcgtcgg gtgatgctgc caacttactg atttagtgta tgatggtgtt
     1621 tttgaggtgc tccagtggct tctgtttcta tcagctgtcc ctcctgttca gctactgacg
     1681 gggtggtgcg taacggcaaa agcaccgccg gacatcagcg ctagcggagt gtatactggc
     1741 ttactatgtt ggcactgatg agggtgtcag tgaagtgctt catgtggcag gagaaaaaag
     1801 gctgcaccgg tgcgtcagca gaatatgtga tacaggatat attccgcttc ctcgctcact
     1861 gactcgctac gctcggtcgt tcgactgcgg cgagcggaaa tggcttacga acggggcgga
     1921 gatttcctgg aagatgccag gaagatactt aacagggaag tgagagggcc gcggcaaagc
     1981 cgtttttcca taggctccgc ccccctgaca agcatcacga aatctgacgc tcaaatcagt
     2041 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tcccctggcg gctccctcgt
     2101 gcgctctcct gttcctgcct ttcggtttac cggtgtcatt ccgctgttat ggccgcgttt
     2161 gtctcattcc acgcctgaca ctcagttccg ggtaggcagt tcgctccaag ctggactgta
     2221 tgcacgaacc ccccgttcag tccgaccgct gcgccttatc cggtaactat cgtcttgagt
     2281 ccaacccgga aagacatgca aaagcaccac tggcagcagc cactggtaat tgatttagag
     2341 gagttagtct tgaagtcatg cgccggttaa ggctaaactg aaaggacaag ttttggtgac
     2401 tgcgctcctc caagccagtt acctcggttc aaagagttgg tagctcagag aaccttcgaa
     2461 aaaccgccct gcaaggcggt tttttcgttt tcagagcaag agattacgcg cagaccaaaa
     2521 cgatctcaag aagatcatct tattaatcag ataaaatatt tctagatttc agtgcaattt
     2581 atctcttcaa atgtagcacc tgaagtcagc cccatacgat ataagttgta attctcatgt
     2641 tagtcatgcc ccgcgcccac cggaaggagc tgactgggtt gaaggctctc aagggcatcg
     2701 gtcgagatcc cggtgcctaa tgagtgagct aacttacatt aattgcgttg cgctcactgc
     2761 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     2821 ggagaggcgg tttgcgtatt gggcgccagg gtggtttttc ttttcaccag tgagacgggc
     2881 aacagctgat tgcccttcac cgcctggccc tgagagagtt gcagcaagcg gtccacgctg
     2941 gtttgcccca gcaggcgaaa atcctgtttg atggtggtta acggcgggat ataacatgag
     3001 ctgtcttcgg tatcgtcgta tcccactacc gagatgtccg caccaacgcg cagcccggac
     3061 tcggtaatgg cgcgcattgc gcccagcgcc atctgatcgt tggcaaccag catcgcagtg
     3121 ggaacgatgc cctcattcag catttgcatg gtttgttgaa aaccggacat ggcactccag
     3181 tcgccttccc gttccgctat cggctgaatt tgattgcgag tgagatattt atgccagcca
     3241 gccagacgca gacgcgccga gacagaactt aatgggcccg ctaacagcgc gatttgctgg
     3301 tgacccaatg cgaccagatg ctccacgccc agtcgcgtac cgtcttcatg ggagaaaata
     3361 atactgttga tgggtgtctg gtcagagaca tcaagaaata acgccggaac attagtgcag
     3421 gcagcttcca cagcaatggc atcctggtca tccagcggat agttaatgat cagcccactg
     3481 acgcgttgcg cgagaagatt gtgcaccgcc gctttacagg cttcgacgcc gcttcgttct
     3541 accatcgaca ccaccacgct ggcacccagt tgatcggcgc gagatttaat cgccgcgaca
     3601 atttgcgacg gcgcgtgcag ggccagactg gaggtggcaa cgccaatcag caacgactgt
     3661 ttgcccgcca gttgttgtgc cacgcggttg ggaatgtaat tcagctccgc catcgccgct
     3721 tccacttttt cccgcgtttt cgcagaaacg tggctggcct ggttcaccac gcgggaaacg
     3781 gtctgataag agacaccggc atactctgcg acatcgtata acgttactgg tttcacattc
     3841 accaccctga attgactctc ttccgggcgc tatcatgcca taccgcgaaa ggttttgcgc
     3901 cattcgatgg tgtccgggat ctcgacgctc tcccttatgc gactcctgca ttaggaaatt
     3961 aatacgactc actat
//
