LOCUS       sequence-244250-        3519 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Slot B, empty (step 1).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3519)
  AUTHORS   Santos-Moreno J, Schaerli Y
  TITLE     A Framework for the Modular and Combinatorial Assembly of Synthetic 
            Gene Circuits.
  JOURNAL   ACS Synth Biol. 2019 Jul 19;8(7):1691-1697. doi: 
            10.1021/acssynbio.9b00174. Epub 2019 Jun 24.
  PUBMED    31185158
REFERENCE   2  (bases 1 to 3519)
  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"; doi: 
            "10.1021/acssynbio.9b00174"; journalName: "ACS Synth Biol"; date: 
            "2019-07-19- 19"; volume: "8"; issue: "7"; pages: "1691-1697"
COMMENT     Sequence Label: pTU-B-000
FEATURES             Location/Qualifiers
     source          1..3519
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(434..452)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      448..495
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      504..590
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      682..709
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     CDS             1212..1403
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(1397..1419)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    1505..1644
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     1560..1579
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(1659..1676)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1830..2418)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1910..1929)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2589..3380,3381..3449))
                     /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     3212..3231
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(3450..3519,1..35))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
       61 gaattcgaaa tccctgaaac tgagactgta gaaaataagc ttgtccagac tattggatcc
      121 aagagatttc tacacgattg agcactgtct cctgcaggct cggtaccaaa ttccagaaaa
      181 gaggcctccc gaaagggggg ccttttttcg ttttggtcct aatagataaa ggataggtct
      241 ggtagtgttg ttcgttctcg caggtaaatc aataatactc agcagttccg tagacttttc
      301 agtgggacag ggtagcgata acagatagat tgtaataaga cacagtaggt gctcgtagtt
      361 gcgtgaagag aaccgctcag gaaatccagt cagaagtatt ggtaatcgtt gaaaactcag
      421 tcgactgctg ccaccgctga gcaataacta gcataacccc ttggggcctc taaacgggtc
      481 ttgaggggtt ttttgccagg catcaaataa aacgaaaggc tcagtcggaa gactgggcct
      541 ttcgttttat ctgttgtttg tcggtgaacg ctctcctgag taggacaaat ccgccgggag
      601 cggatttgaa cgttgtgaag caacggcccg gagggtggcg ggcaggacgc ccgccataaa
      661 ctgccaggca tcaaactaag cagaaggcca tcctgacgga tggccttttt gcgtttcaga
      721 tctcgggcag cgttgggtcc tggccacggg tgcgcatgat cgtgctcctg tcgttgagga
      781 cccggctagg ctggcggggt tgccttactg gttagcagaa tgaatcaccg atacgcgagc
      841 gaacgtgaag cgactgctgc tgcaaaacgt ctgcgacctg agcaacaaca tgaatggtct
      901 tcggtttccg tgtttcgtaa agtctggaaa cgcggaagtc agcgccctgc accattatgt
      961 tccggatctg catcgcagga tgctgctggc taccctgtgg aacacctaca tctgtattaa
     1021 cgaagcgctg gcattgaccc tgagtgattt ttctctggtc ccgccgcatc cataccgcca
     1081 gttgtttacc ctcacaacgt tccagtaacc gggcatgttc atcatcagta acccgtatcg
     1141 tgagcatcct ctctcgtttc atcggtatca ttacccccat gaacagaaat cccccttaca
     1201 cggaggcatc agtgaccaaa caggaaaaaa ccgcccttaa catggcccgc tttatcagaa
     1261 gccagacatt aacgcttctg gagaaactca acgagctgga cgcggatgaa caggcagaca
     1321 tctgtgaatc gcttcacgac cacgctgatg agctttaccg cagctgcctc gcgcgtttcg
     1381 gtgatgacgg tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt
     1441 aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc
     1501 ggggcgcagc catgacccag tcacgtagcg atagcggagt gtatactggc ttaactatgc
     1561 ggcatcagag cagattgtac tgagagtgca ccattgcggt gtgaaatacc gcacagatgc
     1621 gtaaggagaa aataccgcat caggcgctct tccgcttcct cgctcactga ctcgctgcgc
     1681 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     1741 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     1801 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     1861 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     1921 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     1981 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     2041 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     2101 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     2161 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     2221 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt
     2281 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     2341 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     2401 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     2461 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     2521 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     2581 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     2641 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     2701 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     2761 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     2821 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     2881 ttgcgcaacg ttgttgccat tgctgcaggc atcgtggtgt cacgctcgtc gtttggtatg
     2941 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     3001 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     3061 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     3121 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     3181 ccgagttgct cttgcccggc gtcaacacgg gataataccg cgccacatag cagaacttta
     3241 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     3301 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     3361 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     3421 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     3481 tatcagggtt attgtctcat gagcggatac atatttgaa
//
