LOCUS       sequence-244263-        4383 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Slot B, contains sfGFP-MarAn20 (step 1).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4383)
  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 4383)
  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-005
FEATURES             Location/Qualifiers
     source          1..4383
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    166..185
                     /label=Csy4 Site
                     /bound_moiety="Pseudomonas aeruginosa Csy4 (Cas6f)"
                     /note="cleavage site for the Csy4 endoribonuclease (Cermak 
                     et al. 2017)"
     RBS             190..212
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             220..933
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(1298..1316)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1312..1359
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1368..1454
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1546..1573
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     CDS             2076..2267
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(2261..2283)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    2369..2508
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     2424..2443
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(2523..2540)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2694..3282)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2774..2793)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3453..4244,4245..4313))
                     /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     4076..4095
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(4314..4383,1..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 cagcctgcgg tccgggttca ctgccgtata
      181 ggcagtaatt ttgtttaact ttaagaagga gatatacata tgcgtaaagg cgaagaactg
      241 tttaccggtg tggttccgat tctggtggaa ctggacggcg atgttaatgg tcataaattc
      301 agtgttcgcg gcgaaggtga aggcgatgcg acgaacggca aactgaccct gaaatttatc
      361 tgcaccacgg gtaaactgcc ggtcccgtgg ccgacgctgg tgaccacgct gacctatggc
      421 gttcaatgtt ttgcgcgtta cccggatcac atgaaacagc acgacttttt caaatcggcc
      481 atgccggaag gctatgtgca ggaacgtacg attagcttta aagacgatgg tacgtataaa
      541 acccgcgcgg aagtgaaatt cgaaggcgat accctggtta accgtatcga actgaaaggt
      601 atcgatttca aagaagacgg caatattctg ggtcataaac tggaatataa cttcaattcc
      661 cacaacgtgt acatcaccgc ggataaacag aaaaacggca ttaaagccaa tttcaaaatc
      721 cgccataatg tggaagatgg tagcgttcag ctggccgacc actatcagca aaacacgccg
      781 attggtgatg gcccggtcct gctgccggac aatcactacc tgagtaccca gtccgtgctg
      841 tcaaaagatc cgaacgaaaa acgtgaccac atggtcctgc tggaatttgt gacggctgcg
      901 ggtatcaccc acggcatgga cgaactgtat aaaatgtccc gccgtaatac tgacgccatc
      961 acaatccaca gcatcctgga ttggattgaa gactaataat cgctgggacg cccgcctgca
     1021 ggctcggtac caaattccag aaaagaggcc tcccgaaagg ggggcctttt ttcgttttgg
     1081 tcctaataga taaaggatag gtctggtagt gttgttcgtt ctcgcaggta aatcaataat
     1141 actcagcagt tccgtagact tttcagtggg acagggtagc gataacagat agattgtaat
     1201 aagacacagt aggtgctcgt agttgcgtga agagaaccgc tcaggaaatc cagtcagaag
     1261 tattggtaat cgttgaaaac tcagtcgact gctgccaccg ctgagcaata actagcataa
     1321 ccccttgggg cctctaaacg ggtcttgagg ggttttttgc caggcatcaa ataaaacgaa
     1381 aggctcagtc ggaagactgg gcctttcgtt ttatctgttg tttgtcggtg aacgctctcc
     1441 tgagtaggac aaatccgccg ggagcggatt tgaacgttgt gaagcaacgg cccggagggt
     1501 ggcgggcagg acgcccgcca taaactgcca ggcatcaaac taagcagaag gccatcctga
     1561 cggatggcct ttttgcgttt cagatctcgg gcagcgttgg gtcctggcca cgggtgcgca
     1621 tgatcgtgct cctgtcgttg aggacccggc taggctggcg gggttgcctt actggttagc
     1681 agaatgaatc accgatacgc gagcgaacgt gaagcgactg ctgctgcaaa acgtctgcga
     1741 cctgagcaac aacatgaatg gtcttcggtt tccgtgtttc gtaaagtctg gaaacgcgga
     1801 agtcagcgcc ctgcaccatt atgttccgga tctgcatcgc aggatgctgc tggctaccct
     1861 gtggaacacc tacatctgta ttaacgaagc gctggcattg accctgagtg atttttctct
     1921 ggtcccgccg catccatacc gccagttgtt taccctcaca acgttccagt aaccgggcat
     1981 gttcatcatc agtaacccgt atcgtgagca tcctctctcg tttcatcggt atcattaccc
     2041 ccatgaacag aaatccccct tacacggagg catcagtgac caaacaggaa aaaaccgccc
     2101 ttaacatggc ccgctttatc agaagccaga cattaacgct tctggagaaa ctcaacgagc
     2161 tggacgcgga tgaacaggca gacatctgtg aatcgcttca cgaccacgct gatgagcttt
     2221 accgcagctg cctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc
     2281 cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg
     2341 cgtcagcggg tgttggcggg tgtcggggcg cagccatgac ccagtcacgt agcgatagcg
     2401 gagtgtatac tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccattg
     2461 cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg ctcttccgct
     2521 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     2581 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     2641 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     2701 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     2761 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     2821 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     2881 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     2941 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     3001 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     3061 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     3121 ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     3181 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     3241 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     3301 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     3361 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     3421 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     3481 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     3541 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     3601 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     3661 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     3721 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctgc aggcatcgtg
     3781 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     3841 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     3901 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     3961 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     4021 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaac acgggataat
     4081 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     4141 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     4201 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     4261 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     4321 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     4381 gaa
//
