LOCUS       sequence-423497-        5922 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  in vitro transcription.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5922)
  TITLE     Finzi Lab Plasmids 2023
REFERENCE   2  (bases 1 to 5922)
  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: pDME1rrnBT1-T7Te
FEATURES             Location/Qualifiers
     source          1..5922
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             39..1229
                     /codon_start=1
                     /gene="tet"
                     /product="tetracycline efflux protein"
                     /label=TcR
                     /note="confers resistance to tetracycline"
                     /translation="MKSNNALIVILGTVTLDAVGIGLVMPVLPGLLRDIVHSDSIASHY
                     GVLLALYALMQFLCAPVLGALSDRFGRRPVLLASLLGATIDYAIMATTPVLWILYAGRI
                     VAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHAPF
                     LAAAVLNGLNLLLGCFLMQESHKGERRPMPLRAFNPVSSFRWARGMTIVAALMTVFFIM
                     QLVGQVPAALWVIFGEDRFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGEKQAII
                     AGMAADALGYVLLAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSL
                     AALTSLTSIIGPLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSRATST"
     primer_bind     complement(391..410)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             1868..2059
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(2053..2075)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     protein_bind    2993..3009
                     /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)."
     terminator      3605..3676
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3692..3719
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     primer_bind     complement(3877..3894)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4048..4636)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4128..4147)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4807..5598,5599..5667))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     5430..5449
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5668..5772)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5840..5858
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(5885..5890,5891..5907,5908..5913)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
ORIGIN
        1 tatcacagtt aaattgctaa cgcagtcagg caccgtgtat gaaatctaac aatgcgctca
       61 tcgtcatcct cggcaccgtc accctggatg ctgtaggcat aggcttggtt atgccggtac
      121 tgccgggcct cttgcgggat atcgtccatt ccgacagcat cgccagtcac tatggcgtgc
      181 tgctagcgct atatgcgttg atgcaatttc tatgcgcacc cgttctcgga gcactgtccg
      241 accgctttgg ccgccgccca gtcctgctcg cttcgctact tggagccact atcgactacg
      301 cgatcatggc gaccacaccc gtcctgtgga tcctctacgc cggacgcatc gtggccggca
      361 tcaccggcgc cacaggtgcg gttgctggcg cctatatcgc cgacatcacc gatggggaag
      421 atcgggctcg ccacttcggg ctcatgagcg cttgtttcgg cgtgggtatg gtggcaggcc
      481 ccgtggccgg gggactgttg ggcgccatct ccttgcatgc accattcctt gcggcggcgg
      541 tgctcaacgg cctcaaccta ctactgggct gcttcctaat gcaggagtcg cataagggag
      601 agcgtcgacc gatgcccttg agagccttca acccagtcag ctccttccgg tgggcgcggg
      661 gcatgactat cgtcgccgca cttatgactg tcttctttat catgcaactc gtaggacagg
      721 tgccggcagc gctctgggtc attttcggcg aggaccgctt tcgctggagc gcgacgatga
      781 tcggcctgtc gcttgcggta ttcggaatct tgcacgccct cgctcaagcc ttcgtcactg
      841 gtcccgccac caaacgtttc ggcgagaagc aggccattat cgccggcatg gcggccgacg
      901 cgctgggcta cgtcttgctg gcgttcgcga cgcgaggctg gatggccttc cccattatga
      961 ttcttctcgc ttccggcggc atcgggatgc ccgcgttgca ggccatgctg tccaggcagg
     1021 tagatgacga ccatcaggga cagcttcaag gatcgctcgc ggctcttacc agcctaactt
     1081 cgatcattgg accgctgatc gtcacggcga tttatgccgc ctcggcgagc acatggaacg
     1141 ggttggcatg gattgtaggc gccgccctat accttgtctg cctccccgcg ttgcgtcgcg
     1201 gtgcatggag ccgggccacc tcgacctgaa tggaagccgg cggcacctcg ctaacggatt
     1261 caccactcca agaattggag ccaatcaatt cttgcggaga actgtgaatg cgcaaaccaa
     1321 cccttggcag aacgtatcca tcgcgtccgc catctccagc agccgcacgc ggcgcatctc
     1381 gggcagcgtt gggtcctggc cacgggtgcg catgatcgtg ctcctgtcgt tgaggacccg
     1441 gctaggctgg cggggttgcc ttactggtta gcagaatgaa tcaccgatac gcgagcgaac
     1501 gtgaagcgac tgctgctgca aaacgtctgc gacctgagca acaacatgaa tggtcttcgg
     1561 tttccgtgtt tcgtaaagtc tggaaacgcg gaagtcagcg ccctgcacca ttatgttccg
     1621 gatctgcatc gcaggatgct gctggctacc ctgtggaaca cctacatctg tattaacgaa
     1681 gcgctggcat tgaccctgag tgatttttct ctggtcccgc cgcatccata ccgccagttg
     1741 tttaccctca caacgttcca gtaaccgggc atgttcatca tcagtaaccc gtatcgtgag
     1801 catcctctct cgtttcatcg gtatcattac ccccatgaac agaaatcccc cttacacgga
     1861 ggcatcagtg accaaacagg aaaaaaccgc ccttaacatg gcccgcttta tcagaagcca
     1921 gacattaacg cttctggaga aactcaacga gctggacgcg gatgaacagg cagacatctg
     1981 tgaatcgctt cacgaccacg ctgatgagct ttaccgcagc tgcctcgcgc gtttcggtga
     2041 tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc
     2101 ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggca ggtgtcgggg
     2161 cgcagccatg acccagtcac cccatggtgc agtatgaagg cggcggagcc gacaccacgg
     2221 ccaccgatat tatttgcccg atgtacgcgc gcgtggatga agaccagccc ttcccggctt
     2281 tatcaaaaag agtattgact taaagtctaa cctataggat acttacagcg atggagaggt
     2341 gtagtggtaa ccagaagata atatggcttt cgctacctgg agagacgcgc ccgctgatcc
     2401 tttgcgaata cgcccacgcg atgggtaaca gtcttggcgg tttcgctaaa tactggcagg
     2461 cgtttcgtca gtatccccgt ttacagggcg gcttcgtctg ggactgggtg gatcagtcgc
     2521 tgattaaata tgatgaaaac ggcaacccgt ggtaccggct tacggcggtg atttttgcga
     2581 tacgccgaac gtcgatcgag ctcaacggaa ttctcgccag ttctgtatga acggtctggt
     2641 ctttgccgac cgcacgtcgc atccagcgct gacggaagca aaacaccagc agcagttttt
     2701 cagttccgtt tatccgggca aaccatcgaa gtgaccagcg aatacctgtt ccgtcatagc
     2761 gataacgagc tcctgcactg gatggtggcg ctggatggta agccgctggc aagcggtgaa
     2821 gtgcctctgg atgtcgctcc acaaggtaaa cagttgattg aactgtctga actaccgcag
     2881 ccggagagcg ccgggcaact ctggctcaca gtacgcgtag tgcaaccgaa cgcgaccgca
     2941 tggtcagaac tagatgtaca gaagtcagat tagcagtagc gtctggcgga aattgtgagc
     3001 ggataacaat taaacctcag tgtgacgctc cccgccgcgt cccacgccat cccgcatctg
     3061 accaccagcg aaatggattt ttgcactcga gctgggtaat aagcgttggc cccgacaact
     3121 ccatctgctt cttccgtgcg agatgtggat tggcgataaa aaacaactgc tgacgccgct
     3181 gcgcgatcag ttcacccgtg caccgctgga taacgacatt ggcgtaagtg aagcgacccg
     3241 cattgaccct aacgcctggg tcgaacactg gaaggcggcg ggccattacc aggccgaagc
     3301 agcgcgccgc ttgcgatttc gaccaacagc tgctgatgcg gtgctgatta cgaccgctca
     3361 cgcgtggcag catcagggga aaaccttatt tatcagccgg aagcaccgtg taacttccag
     3421 gacatctatg atggcgatta ccgttgatgt tgaagtggcg agcgatacac cgcatccggc
     3481 gcggatttct gaaaagatgt cgtgctactc catactcggg gtcctgtatg ggctcggatt
     3541 agcggccgca agaaaactat cccgaccgcc ttactgccgc ctgtttgcta ctagagccag
     3601 gcatcaaata aaacgaaagg ctcagtcgaa agactgggcc tttcgtttta tctgttgttt
     3661 gtcggtgaac gctctctact agagtcacac tggctcacct tcgggtgggc ctttctgcgt
     3721 ttatatgcgt ttatagatct acagaagcgc gattgtcggg cacaggctta actatgcggc
     3781 atcagagcag acagagtacg acgatttgcg atggaacttg gagataccgc acagatgcgt
     3841 aaggagaaaa taccgcatca ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     3901 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     3961 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     4021 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     4081 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     4141 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     4201 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     4261 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     4321 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     4381 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     4441 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg
     4501 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     4561 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     4621 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     4681 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     4741 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     4801 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     4861 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     4921 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     4981 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     5041 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     5101 gcgcaacgtt gttgccattg ctgcaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     5161 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     5221 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     5281 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     5341 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     5401 gagttgctct tgcccggcgt caacacggga taataccgcg ccacatagca gaactttaaa
     5461 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     5521 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     5581 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     5641 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     5701 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     5761 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat
     5821 catgacatta acctataaaa ataggcgtat cacgaggccc tttcgtcttc aagaattctc
     5881 atgtttgaca gcttatcatc gataagcttt aatgcggtag tt
//
