LOCUS       sequence-244249-        4405 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Slot C, contains RepA70-mKate2 (step 1).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4405)
  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 4405)
  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-C-005
FEATURES             Location/Qualifiers
     source          1..4405
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    164..183
                     /label=Csy4 Site
                     /bound_moiety="Pseudomonas aeruginosa Csy4 (Cas6f)"
                     /note="cleavage site for the Csy4 endoribonuclease (Cermak 
                     et al. 2017)"
     RBS             188..210
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(1320..1338)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1334..1381
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1390..1476
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1568..1595
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     CDS             2098..2289
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(2283..2305)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    2391..2530
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     2446..2465
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(2545..2562)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2716..3304)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2796..2815)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3475..4266,4267..4335))
                     /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     4098..4117
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(4336..4405,1..35))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
       61 gaattcagaa gtattggtaa tcgttgaaaa ctcagtcgac gcacttactg aagacgtcct
      121 attacactcg tcgttggaaa ctgaagatca gcctgcggtc cgggttcact gccgtatagg
      181 cagtaatttt gtttaacttt aagaaggaga tatacatatg aatcagtcat tcatctcgga
      241 catcttatat gccgacatcg aatcgaaggc taaggaactt acagtcaatt ccaacaatac
      301 tgtccagccg gtcgcgctta tgcgcttagg agttttcgtt cccaaacctt ccaagagcaa
      361 aggagaaagt aaggaaattg acgccaccaa agccttctct caactggaga ttgctaaagc
      421 agagggcatg gttagtaaag gagaagaaaa taacatggca ctgattaagg agaacatgca
      481 catgaagctg tacatggagg gcaccgtgaa caaccaccac ttcaagtgca catccgaggg
      541 cgaaggcaag ccctacgagg gcacccagac catgagaatc aaggccgtcg agggcggccc
      601 tctccccttc gccttcgaca tcctggctac cagcttcatg tacggcagca aaaccttcat
      661 caaccacacc cagggcatcc ccgacttctt taagcagtcc ttccctgagg gcttcacatg
      721 ggagagagtc accacatacg aagacggggg cgtgctgacc gctacccagg acaccagcct
      781 ccaggacggc tgcctcatct acaacgtcaa gatcagaggg gtgaacttcc catccaacgg
      841 ccctgtgatg cagaagaaaa cactcggctg ggaggcctcc accgagaccc tgtaccccgc
      901 tgacggcggc ctggaaggca gagccgacat ggccctgaag ctcgtgggcg ggggccacct
      961 gatctgcaac ttgaagacca catacagatc caagaaaccc gctaagaacc tcaagatgcc
     1021 cggcgtctac tatgtggaca gaagactgga aagaatcaag gaggccgaca aagagaccta
     1081 cgtcgagcag cacgaggtgg ctgtggccag atactgcgac ctccctagca aactggggca
     1141 cagagtctaa taatcgctgg gacgcccggc ggccgcggaa acacagaaaa aagcccgcac
     1201 ctgacagtgc gggctttttt tttcgaccaa aggtagcgaa cgacgagtca ctgttgagga
     1261 taaatacttt ctctactagg cgcctgttac acaggtcctc agcggcgcgc ctgctgccac
     1321 cgctgagcaa taactagcat aaccccttgg ggcctctaaa cgggtcttga ggggtttttt
     1381 gccaggcatc aaataaaacg aaaggctcag tcggaagact gggcctttcg ttttatctgt
     1441 tgtttgtcgg tgaacgctct cctgagtagg acaaatccgc cgggagcgga tttgaacgtt
     1501 gtgaagcaac ggcccggagg gtggcgggca ggacgcccgc cataaactgc caggcatcaa
     1561 actaagcaga aggccatcct gacggatggc ctttttgcgt ttcagatctc gggcagcgtt
     1621 gggtcctggc cacgggtgcg catgatcgtg ctcctgtcgt tgaggacccg gctaggctgg
     1681 cggggttgcc ttactggtta gcagaatgaa tcaccgatac gcgagcgaac gtgaagcgac
     1741 tgctgctgca aaacgtctgc gacctgagca acaacatgaa tggtcttcgg tttccgtgtt
     1801 tcgtaaagtc tggaaacgcg gaagtcagcg ccctgcacca ttatgttccg gatctgcatc
     1861 gcaggatgct gctggctacc ctgtggaaca cctacatctg tattaacgaa gcgctggcat
     1921 tgaccctgag tgatttttct ctggtcccgc cgcatccata ccgccagttg tttaccctca
     1981 caacgttcca gtaaccgggc atgttcatca tcagtaaccc gtatcgtgag catcctctct
     2041 cgtttcatcg gtatcattac ccccatgaac agaaatcccc cttacacgga ggcatcagtg
     2101 accaaacagg aaaaaaccgc ccttaacatg gcccgcttta tcagaagcca gacattaacg
     2161 cttctggaga aactcaacga gctggacgcg gatgaacagg cagacatctg tgaatcgctt
     2221 cacgaccacg ctgatgagct ttaccgcagc tgcctcgcgc gtttcggtga tgacggtgaa
     2281 aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccggg
     2341 agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg cgcagccatg
     2401 acccagtcac gtagcgatag cggagtgtat actggcttaa ctatgcggca tcagagcaga
     2461 ttgtactgag agtgcaccat tgcggtgtga aataccgcac agatgcgtaa ggagaaaata
     2521 ccgcatcagg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     2581 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     2641 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     2701 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     2761 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     2821 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     2881 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     2941 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     3001 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     3061 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     3121 cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct
     3181 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     3241 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     3301 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     3361 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     3421 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     3481 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     3541 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     3601 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     3661 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     3721 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     3781 tgccattgct gcaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     3841 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     3901 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     3961 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     4021 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     4081 cccggcgtca acacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     4141 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     4201 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     4261 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     4321 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     4381 tctcatgagc ggatacatat ttgaa
//
