LOCUS       sequence-45110-e        7428 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 7428)
  AUTHORS   Seeliger JC, Topp S, Sogi KM, Previti ML, Gallivan JP, Bertozzi CR
  TITLE     A riboswitch-based inducible gene expression system for 
            mycobacteria.
  JOURNAL   PLoS One. 2012;7(1):e29266. Epub 2012 Jan 18.
  PUBMED    22279533
REFERENCE   2  (bases 1 to 7428)
  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: "PLoS One.";
            date: "2012"; volume: "7(1)"; pages: "e29266. Epub 2012 Jan 18"
COMMENT     Sequence Label: pST5832
FEATURES             Location/Qualifiers
     source          1..7428
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             2197..5271
                     /codon_start=1
                     /gene="lacZ"
                     /product="beta-galactosidase"
                     /label=lacZ
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVNSAFHLWCNGRWVGYG
                     QDSRLPSEFDLSAFLRAGENRLAVMVLRWSDGSYLEDQDMWRMSGIFRDVSLLHKPTTQ
                     ISDFHVATRFNDDFSRAVLEAEVQMCGELRDYLRVTVSLWQGETQVASGTAPFGGEIID
                     ERGGYADRVTLRLNVENPKLWSAEIPNLYRAVVELHTADGTLIEAEACDVGFREVRIEN
                     GLLLLNGKPLLIRGVNRHEHHPLHGQVMDEQTMVQDILLMKQNNFNAVRCSHYPNHPLW
                     YTLCDRYGLYVVDEANIETHGMVPMNRLTDDPRWLPAMSERVTRMVQRDRNHPSVIIWS
                     LGNESGHGANHDALYRWIKSVDPSRPVQYEGGGADTTATDIICPMYARVDEDQPFPAVP
                     KWSIKKWLSLPGETRPLILCEYAHAMGNSLGGFAKYWQAFRQYPRLQGGFVWDWVDQSL
                     IKYDENGNPWSAYGGDFGDTPNDRQFCMNGLVFADRTPHPALTEAKHQQQFFQFRLSGQ
                     TIEVTSEYLFRHSDNELLHWMVALDGKPLASGEVPLDVAPQGKQLIELPELPQPESAGQ
                     LWLTVRVVQPNATAWSEAGHISAWQQWRLAENLSVTLPAASHAIPHLTTSEMDFCIELG
                     NKRWQFNRQSGFLSQMWIGDKKQLLTPLRDQFTRAPLDNDIGVSEATRIDPNAWVERWK
                     AAGHYQAEAALLQCTADTLADAVLITTAHAWQHQGKTLFISRKTYRIDGSGQMAITVDV
                     EVASDTPHPARIGLNCQLAQVAERVNWLGLGPQENYPDRLTAACFDRWDLPLSDMYTPY
                     VFPSENGLRCGTRELNYGPHQWRGDFQFNISRYSQQQLMETSHRHLLHAEEGTWLNIDG
                     FHMGIGGDDSWSPSVSAEFQLSAGRYHYQLVWCQK"
     primer_bind     complement(2217..2234)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2217..2233)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2226..2248)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     terminator      5327..5373
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(5523..5542)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             5534..6349
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPHAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHNLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(5607..5626)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      6680..7268
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7169..7188
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gagcccacca gctccgtaag ttcgggcgct gtgtggctcg tacccgcgca ttcaggcggc
       61 agggggtcta acgggtctaa ggcggcgtgt acggccgcca cagcggctct cagcggcccg
      121 gaaacgtcct cgaaacgacg catgtgttcc tcctggttgg tacaggtggt tgggggtgct
      181 cggctgtcgc tggtgttcca ccaccagggc tcgacgggag agcgggggag tgtgcagttg
      241 tggggtggcc cctcagcgaa atatctgact tggagctcgt gtcggaccat acaccggtga
      301 ttaatcgtgg tctactacca agcgtgagcc acgtcgccga cgaatttgag cagctctggc
      361 tgccgtactg gccgctggca agcgacgatc tgctcgaggg gatctaccgc caaagccgcg
      421 cgtcggccct aggccgccgg tacatcgagg cgaacccaac agcgctggca aacctgctgg
      481 tcgtggacgt agaccatcca gacgcagcgc tccgagcgct cagcgcccgg gggtcccatc
      541 cgctgcccaa cgcgatcgtg ggcaatcgcg ccaacggcca cgcacacgca gtgtgggcac
      601 tcaacgcccc tgttccacgc accgaatacg cgcggcgtaa gccgctcgca tacatggcgg
      661 cgtgcgccga aggccttcgg cgcgccgtcg atggcgaccg cagttactca ggcctcatga
      721 ccaaaaaccc cggccacatc gcctgggaaa cggaatggct ccactcagat ctctacacac
      781 tcagccacat cgaggccgag ctcggcgcga acatgccacc gccgcgctgg cgtcagcaga
      841 ccacgtacaa agcggctccg acgccgctag ggcggaattg cgcactgttc gattccgtca
      901 ggttgtgggc ctatcgtccc gccctcatgc ggatctacct gccgacccgg aacgtggacg
      961 gactcggccg cgcgatctat gccgagtgcc acgcgcgaaa cgccgaattt ccgtgcaacg
     1021 acgtgtgtcc cggaccgcta ccggacagcg aggtccgcgc catcgccaac agcatttggc
     1081 gttggatcac aaccaagtcg cgcatttggg cggacgggat cgtggtctac gaggccacac
     1141 tcagtgcgcg ccagtcggcc atctcgcgga agggcgcagc agcgcgcacg gcggcgagca
     1201 cagttgcgcg gcgcgcaaag tccgcgtcag ccatggaggc attgctatga gcgacggcta
     1261 cagcgacggc tacaaccggc agccgactgt ccgcaaaaag cggcgcgtga ccgccgccga
     1321 aggcgctcga atcaccggac tatccgaacg ccacgtcgtc cggctcgtgg cgcaggaacg
     1381 cagcgagtgg ctcgccgagc aggctgcacg ccgcgaacgc atccgcgcct atcacgacga
     1441 cgagggccac tcttggccgc aaacggccaa acatttcggg ctgcatctgg acaccgttaa
     1501 gcgactcggc tatcgggcga ggaaagagcg tgcggcagaa caggaagcgg ctcaaaaggc
     1561 ccacaacgaa gccgacaatc caccgctgtt ctaacgcaat tggggagcgg gtgtcgcggg
     1621 ggttccgtgg ggggttccgt tgcaacgggt cggacaggta aaagtcctgg tagacgctag
     1681 ttttctggtt tgggccatgc ctgtctcgtt gcgtgtttcg ttgcgcccgt tttgaatacc
     1741 agccagacga gacggggttc tacgaatctt ggtcgatacc aagccatttc cgctgaatat
     1801 cggggagctc accgccagaa tcggtggttg tggtgatgta cgtggcgaac tccgttgtag
     1861 tgcctgtggt ggcatccgtg gcgcggccgc ggtaccagat ctttaaatct agaggtgacc
     1921 acaacgcgcc cgctttgatc ggggacgtct gcggccgacc atttacgggt cttgttgtcg
     1981 ttggcggtca tgggccgaac atactcaccc ggatcggagg gccgaggaca aggtcgaacg
     2041 aggggcatga cccggtgcgg ggcttcttgc actcggcata ggcgagtgct aagaataacg
     2101 ttggcactcg cgaccgggat acgactcact ataggtaccg gtgataccag catcgtcttg
     2161 atgcccttgg cagcaccctg ctaaggaggc aacaagatga ccatgattac ggattcactg
     2221 gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg ttacccaact taatcgcctt
     2281 gcagcacatc cccctttcgc cagctggcgt aatagcgaag aggcccgcac cgatcgccct
     2341 tcccaacagt tgcgcagcct gaatggcgaa tggcgctttg cctggtttcc ggcaccagaa
     2401 gcggtgccgg aaagctggct ggagtgcgat cttcctgagg ccgatactgt cgtcgtcccc
     2461 tcaaactggc agatgcacgg ttacgatgcg cccatctaca ccaacgtaac ctatcccatt
     2521 acggtcaatc cgccgtttgt tcccacggag aatccgacgg gttgttactc gctcacattt
     2581 aatgttgatg aaagctggct acaggaaggc cagacgcgaa ttatttttga tggcgttaac
     2641 tcggcgtttc atctgtggtg caacgggcgc tgggtcggtt acggccagga cagtcgtttg
     2701 ccgtctgaat ttgacctgag cgcattttta cgcgccggag aaaaccgcct cgcggtgatg
     2761 gtgctgcgtt ggagtgacgg cagttatctg gaagatcagg atatgtggcg gatgagcggc
     2821 attttccgtg acgtctcgtt gctgcataaa ccgactacac aaatcagcga tttccatgtt
     2881 gccactcgct ttaatgatga tttcagccgc gctgtactgg aggctgaagt tcagatgtgc
     2941 ggcgagttgc gtgactacct acgggtaaca gtttctttat ggcagggtga aacgcaggtc
     3001 gccagcggca ccgcgccttt cggcggtgaa attatcgatg agcgtggtgg ttatgccgat
     3061 cgcgtcacac tacgtctgaa cgtcgaaaac ccgaaactgt ggagcgccga aatcccgaat
     3121 ctctatcgtg cggtggttga actgcacacc gccgacggca cgctgattga agcagaagcc
     3181 tgcgatgtcg gtttccgcga ggtgcggatt gaaaatggtc tgctgctgct gaacggcaag
     3241 ccgttgctga ttcgaggcgt taaccgtcac gagcatcatc ctctgcatgg tcaggtcatg
     3301 gatgagcaga cgatggtgca ggatatcctg ctgatgaagc agaacaactt taacgccgtg
     3361 cgctgttcgc attatccgaa ccatccgctg tggtacacgc tgtgcgaccg ctacggcctg
     3421 tatgtggtgg atgaagccaa tattgaaacc cacggcatgg tgccaatgaa tcgtctgacc
     3481 gatgatccgc gctggctacc ggcgatgagc gaacgcgtaa cgcgaatggt gcagcgcgat
     3541 cgtaatcacc cgagtgtgat catctggtcg ctggggaatg aatcaggcca cggcgctaat
     3601 cacgacgcgc tgtatcgctg gatcaaatct gtcgatcctt cccgcccggt gcagtatgaa
     3661 ggcggcggag ccgacaccac ggccaccgat attatttgcc cgatgtacgc gcgcgtggat
     3721 gaagaccagc ccttcccggc tgtgccgaaa tggtccatca aaaaatggct ttcgctacct
     3781 ggagagacgc gcccgctgat cctttgcgaa tacgcccacg cgatgggtaa cagtcttggc
     3841 ggtttcgcta aatactggca ggcgtttcgt cagtatcccc gtttacaggg cggcttcgtc
     3901 tgggactggg tggatcagtc gctgattaaa tatgatgaaa acggcaaccc gtggtcggct
     3961 tacggcggtg attttggcga tacgccgaac gatcgccagt tctgtatgaa cggtctggtc
     4021 tttgccgacc gcacgccgca tccagcgctg acggaagcaa aacaccagca gcagtttttc
     4081 cagttccgtt tatccgggca aaccatcgaa gtgaccagcg aatacctgtt ccgtcatagc
     4141 gataacgagc tcctgcactg gatggtggcg ctggatggta agccgctggc aagcggtgaa
     4201 gtgcctctgg atgtcgctcc acaaggtaaa cagttgattg aactgcctga actaccgcag
     4261 ccggagagcg ccgggcaact ctggctcaca gtacgcgtag tgcaaccgaa cgcgaccgca
     4321 tggtcagaag ccgggcacat cagcgcctgg cagcagtggc gtctggcgga aaacctcagt
     4381 gtgacgctcc ccgccgcgtc ccacgccatc ccgcatctga ccaccagcga aatggatttt
     4441 tgcatcgagc tgggtaataa gcgttggcaa tttaaccgcc agtcaggctt tctttcacag
     4501 atgtggattg gcgataaaaa acaactgctg acgccgctgc gcgatcagtt cacccgtgca
     4561 ccgctggata acgacattgg cgtaagtgaa gcgacccgca ttgaccctaa cgcctgggtc
     4621 gaacgctgga aggcggcggg ccattaccag gccgaagcag cgttgttgca gtgcacggca
     4681 gatacacttg ctgatgcggt gctgattacg accgctcacg cgtggcagca tcaggggaaa
     4741 accttattta tcagccggaa aacctaccgg attgatggta gtggtcaaat ggcgattacc
     4801 gttgatgttg aagtggcgag cgatacaccg catccggcgc ggattggcct gaactgccag
     4861 ctggcgcagg tagcagagcg ggtaaactgg ctcggattag ggccgcaaga aaactatccc
     4921 gaccgcctta ctgccgcctg ttttgaccgc tgggatctgc cattgtcaga catgtatacc
     4981 ccgtacgtct tcccgagcga aaacggtctg cgctgcggga cgcgcgaatt gaattatggc
     5041 ccacaccagt ggcgcggcga cttccagttc aacatcagcc gctacagtca acagcaactg
     5101 atggaaacca gccatcgcca tctgctgcac gcggaagaag gcacatggct gaatatcgac
     5161 ggtttccata tggggattgg tggcgacgac tcctggagcc cgtcagtatc ggcggaattc
     5221 cagctgagcg ccggtcgcta ccattaccag ttggtctggt gtcaaaaata ataaaagctt
     5281 atcgatgtcg acgtagttaa ctagcgtacg atcgactgcc aggcatcaaa taaaacgaaa
     5341 ggctcagtcg aaagactggg cctttcgttt tatgccatca tggccgcggt gatcagctag
     5401 ccacctgacg tcgggggggg gggaaagcca cgttgtgtct caaaatctct gatgttacat
     5461 tgcacaagat aaaaatatat catcatgaac aataaaactg tctgcttaca taaacagtaa
     5521 tacaaggggt gttatgagcc atattcaacg ggaaacgtct tgctcgaggc cgcgattaaa
     5581 ttccaacatg gatgctgatt tatatgggta taaatgggct cgcgataatg tcgggcaatc
     5641 aggtgcgaca atctatcgct tgtatgggaa gccccatgcg ccagagttgt ttctgaaaca
     5701 tggcaaaggt agcgttgcca atgatgttac agatgagatg gtcagactaa actggctgac
     5761 ggaatttatg cctcttccga ccatcaagca ttttatccgt actcctgatg atgcatggtt
     5821 actcaccact gcgatccccg ggaaaacagc attccaggta ttagaagaat atcctgattc
     5881 aggtgaaaat attgttgatg cgctggcagt gttcctgcgc cggttgcatt cgattcctgt
     5941 ttgtaattgt ccttttaaca gcgatcgcgt atttcgtctc gctcaggcgc aatcacgaat
     6001 gaataacggt ttggttgatg cgagtgattt tgatgacgag cgtaatggct ggcctgttga
     6061 acaagtctgg aaagaaatgc ataatctttt gccattctca ccggattcag tcgtcactca
     6121 tggtgatttc tcacttgata accttatttt tgacgagggg aaattaatag gttgtattga
     6181 tgttggacga gtcggaatcg cagaccgata ccaggatctt gccatcctat ggaactgcct
     6241 cggtgagttt tctccttcat tacagaaacg gctttttcaa aaatatggta ttgataatcc
     6301 tgatatgaat aaattgcagt ttcatttgat gctcgatgag tttttctaat cagaattggt
     6361 taattggttg taacactggc agagcattac gctgacttga cgggacggcg gctttgttga
     6421 ataaatcgaa cttttgctga gttgaaggat cagatcacgc atcttcccga caacgcagac
     6481 cgttccgtgg caaagcaaaa gttcaaaatc accaactggt ccacctacaa caaagctctc
     6541 atcaaccgtg gctccctcac tttctggctg gatgatgggg cgattcaggc ctggtatgag
     6601 tcagcaacac cttcttcacg aggcagacct cactagttcc atgagcgtca gaccccgtag
     6661 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     6721 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     6781 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc
     6841 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     6901 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     6961 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     7021 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     7081 gcgccacgct tcccgagggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     7141 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     7201 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     7261 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     7321 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg
     7381 agtgagctga taccgctcgc cgcagccgaa cgaccgagcg caacgcgt
//
