LOCUS       sequence-12384-e        6498 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 6498)
  TITLE     pMOTag2T
REFERENCE   2  (bases 1 to 6498)
  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"
COMMENT     Sequence Label: pUB39
FEATURES             Location/Qualifiers
     source          1..6498
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     720..739
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        720..738
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    745..763
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     RBS             2299..2307
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     CDS             2969..3343
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     terminator      3746..3793
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      3879..3926
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(4043..4062)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(4273..4292)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4392..4414
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4452..4470)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4538..4642
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4643..4711,4712..5503)
                     /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"
     primer_bind     complement(4861..4880)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5674..6262
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6163..6182
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6416..6433
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gaattcgagc tcatatagtt ggtatgtatt ctaattccag actactggcg tggataaaca
       61 tgtcccctga ttaaaaggaa agattccata gcccattagt gcaaagataa ttggtacacc
      121 gtcaaaaaca catgggagac cacacgatac gacactgtga ccgtagcatc aattccgcac
      181 tcgatataac tctatcgaag aacttccatg gtacaaactg gtcgccacgg tctgctccac
      241 acggagatca tcgtatcatt tttatcgata gcggccgcta tcgatgtatg ccttggccct
      301 gatggcatgc caatttcact acaacggact atgtggaccc cgttatcatg gaaatgcgct
      361 agttggagga agttagaccg cgccggaaaa gagaggggta gagaaaatga caacttggaa
      421 gatatccaca cacgcacggt gaaacgttag caacaattat tagggaagcg acgcttgccc
      481 tagtcccacg agtaaccaag actccaaaag cctttctggc acagagagcg agccgaaatg
      541 gaaaagagaa acaatgcctg cactaacact actgagcgat tcgcctcgcc gcggaggacc
      601 gaatactaat aacgacactt gcggtcaaaa agtagaagaa caaatgctca acgatgagtg
      661 aatcaggtta gggtagttgg aaaattatac agaatgtctt tggcaacaca ccggctgatt
      721 aatacgactc actataggga gatctcccta tcagtgatag agatctcccg ggctgcacgc
      781 gccttcgagt tttttttcct tttccccatt tttttcaact tgaagacttc aattacacca
      841 aaaagtaaaa ttcacaagct aattccagct gtcgacgcgg ggtttctgta ctatattgca
      901 gcaaaagact agaagcaagc agcgcatata gcgcaaacaa tcggggtttc aacaaaaacg
      961 ggagcgactc acaaagctta tggactgcca tacaaaggag acactagggg tcacacaatg
     1021 gaggcgatca acgatgctaa cactatcact gctttacgcc atcactccag cggacggcgc
     1081 caaagaagcc cttgaataca aaacttggac aaaccactgc ggactggcgg ccacactgag
     1141 aaaggttgcc ggtggagtat taacgaaact gaaaagccac attagctacc ggaaaaaact
     1201 ggaagaaatg gaaacgaagc tacgaatcta cgcactaaaa ggagacggag tgggagagca
     1261 aaaatcagcg gagatactag caacaacggc agccctaatg cgacaaaaag cactcacacc
     1321 agaagaagca aatttgaaaa cagcgctgaa ggcggcagga ttcgcaggcg aaggagcggc
     1381 agccgtcagc agctacctga tgacactcgg gacactgaca acaagcggat ctgcgcactg
     1441 cctaagcaac gaaggcggcg acggtgacgg aaaagaccaa cttgcgccga aaggctgccg
     1501 gcacggcaca gaagcagact tcgacgcagg agccggcccg gcagaatctg aagtagccga
     1561 cagcggcttc gcgcaagtac caggcaaaca ggacggagca aacgcaggcc aagcaaacat
     1621 gtgcgcattg ttcacacacc aagcaacgcc gcacagctca cagggcatat acataaccgg
     1681 ggcacagaca aaaccttcat tcgggtacgg catgctgaca atcggcacga cggaccagac
     1741 catcggcttg aaactttcgg acataaaggg caaacaagca gacagcgcgc agaaattctg
     1801 gagcagctgc cacgcagcag tcaaagccgc ccaagatatg aaggcagacc cagccctaaa
     1861 agtcgaccag acgctcctag ctgttcttgt ggcttctccg gagatggctg aaatactgaa
     1921 actagaagcg gcagcatcac agcaaaaagg accagaggaa gtgacgatcg acctagccac
     1981 cgagaaaaac aattatttcg gaaccaacaa caacaaacta gagccgctct ggactaaaat
     2041 caaaggacag aatatagttg acttggcggc gaccaaaggc agcacgaaag agttaggaac
     2101 agtcacagac acggccgagc tacaaaaact tttaagttat tattacacgg tcaacaaaga
     2161 agaacagaaa aaaacagcgg agaaaataac taaactcgaa accgaactag cagatcaaaa
     2221 aggcaaatcc cctgaaagcg agtgcaataa aatatctgag gaacccaaat gcaacgagga
     2281 caagatatgc agttggcata aggaggttaa agcgggagaa aagcactgca aatttaactc
     2341 aacaaaagca aaagaaaagg gggtctctgt aacacaaact caaactgcag gaggaaccga
     2401 agcgacaaca gataaatgca aagggaaatt ggaagatacc tgcaagaagg agagcaactg
     2461 caaatgggaa aataatgctt gcaaagattc ctctattcta gtaaccaaga aattcgccct
     2521 caccgtggtt tctgctgcat ttgtggcctt gcttttttaa ggatcctttt ccccctcttt
     2581 ttcttaaaaa ttcttgctac ttgaaaactc ctgatatatt ttaacacgca aattacccga
     2641 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaacccg cgtcgacagc tggaattgcc
     2701 catttagttg gcttttccct tgtctcgtgt cttttccgtg gaaaggttcc cggagtaatc
     2761 tgatggcaca gcagggaggt gcgcctgcag gttggttagg aaggggggat gatgtaaaag
     2821 aagaaaatgg ggggattcga gcccgggcac agcaaggtct tctgaaattc atgttttttt
     2881 tttttttact ctgcattgca gtctccgctc ttatttagtt ttgctttacg taaggtctcg
     2941 ttgctgccat aaaataagct atatcaccat ggccaagttg accagtgccg ttccggtgct
     3001 caccgcgcgc gacgtcgccg gagcggtcga gttctggacc gaccggctcg ggttctcccg
     3061 ggacttcgtg gaggacgact tcgccggtgt ggtccgggac gacgtgaccc tgttcatcag
     3121 cgcggtccag gaccaggtgg tgccggacaa caccctggcc tgggtgtggg tgcgcggcct
     3181 ggacgagctg tacgccgagt ggtcggaggt cgtgtccacg aacttccggg acgcctccgg
     3241 gccggccatg accgagatcg gcgagcagcc gtgggggcgg gagttcgccc tgcgcgaccc
     3301 ggccggcaac tgcgtgcact tcgtggccga ggagcaggac tgaccgacgc cgaccaacac
     3361 cgccggtccg acggcggccc acgggtccca ggcctcggag atcctaacac cgggttgtgt
     3421 ggccaaaatt gttctgtagt cgctgtgagt tgacacggct agtgcttatg attttcctcg
     3481 cgtgtggtgc ctgtactcag ccctatgcct tatttgcaac acatttacgt acagcgcaca
     3541 agagaagaga agatcacttg aagataataa atatagggtt gtaggcatct tgtttaactc
     3601 aaattttctc gtcttggtgt gtcgacatga ttgaaatagt gccaccagtt gtgtttgatg
     3661 cgtttgttat ctatgcagta ttctgcaggc tgctaacaaa gcccgaaagg aagctgagtt
     3721 ggctgctgcc accgctgagc aataactagc ataacccctt ggggcctcta aacgggtctt
     3781 gaggggtttt ttgctgaaag gaggaactat atccggatag ggctgctaac aaagcccgaa
     3841 aggaagctga gttggctgct gccaccgctg agcaataact agcataaccc cttggggcct
     3901 ctaaacgggt cttgaggggt tttttgctga aaggaggaac tatatccgga tagggcccct
     3961 gcaggcatgc aagctagcca gatcctctac gccggacgca tcgtggccgg catcaccggc
     4021 gccacaggtg cggttgctgg cgcctatatc gccgacatca ccgatgggga agatcgggct
     4081 cgccacttcg ggctcatgag cgcttgtttc ggcgtgggta tggtggcagg ccccgtggcc
     4141 gggggactgt tgggcgccat ctccttgcac cattccttgc ggcggcggtg ctcaacggcc
     4201 tcaacctact actgggctgc ttcctaatgc aggagtcgca taagggagag cgtcgatatg
     4261 gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagcccc gacacccgcc
     4321 aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc
     4381 tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc
     4441 gagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt
     4501 ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     4561 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     4621 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     4681 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     4741 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
     4801 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
     4861 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
     4921 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     4981 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     5041 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
     5101 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     5161 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac
     5221 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     5281 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     5341 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     5401 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     5461 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
     5521 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     5581 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     5641 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     5701 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     5761 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     5821 ccttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     5881 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     5941 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     6001 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     6061 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     6121 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     6181 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     6241 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     6301 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
     6361 tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga
     6421 gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg
     6481 gccgattcat taatgcag
//
