LOCUS       sequence-460511-        5535 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid carries copper sensor circuit genes for whole-cell sensing 
            of copper ions with RBS30.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5535)
  AUTHORS   Koksaldi IC, Avci E, Kose S, Ozkul G, Kehribar ES, Safak Seker UO
  TITLE     Genetically engineered bacterial biofilm materials enhances portable
            whole cell sensing.
  JOURNAL   Biosens Bioelectron. 2024 Aug 10;264:116644. doi: 
            10.1016/j.bios.2024.116644.
  PUBMED    39137519
REFERENCE   2  (bases 1 to 5535)
  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"; journalName: "Biosens 
            Bioelectron."; date: "2024-08-10"; volume: "264:116644. doi"; pages:
            " 10.1016/j.bios.2024.116644"
COMMENT     Sequence Label: pET-22b(+) pCopA RBS30 sfgfp
FEATURES             Location/Qualifiers
     source          1..5535
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             439..630
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(624..646)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    732..871
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     787..806
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(886..903)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1057..1645)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1137..1156)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1816..2607,2608..2676))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     2439..2458
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2677..2781)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2807..3262)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2944..2965)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3156..3175
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     terminator      complement(3299..3346)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     3342..3360
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(3413..3430)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(3440..4153)
                     /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"
     RBS             4160..4171
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
ORIGIN
        1 ttgaggaccc ggctaggctg gcggggttgc cttactggtt agcagaatga atcaccgata
       61 cgcgagcgaa cgtgaagcga ctgctgctgc aaaacgtctg cgacctgagc aacaacatga
      121 atggtcttcg gtttccgtgt ttcgtaaagt ctggaaacgc ggaagtcagc gccctgcacc
      181 attatgttcc ggatctgcat cgcaggatgc tgctggctac cctgtggaac acctacatct
      241 gtattaacga agcgctggca ttgaccctga gtgatttttc tctggtcccg ccgcatccat
      301 accgccagtt gtttaccctc acaacgttcc agtaaccggg catgttcatc atcagtaacc
      361 cgtatcgtga gcatcctctc tcgtttcatc ggtatcatta cccccatgaa cagaaatccc
      421 ccttacacgg aggcatcagt gaccaaacag gaaaaaaccg cccttaacat ggcccgcttt
      481 atcagaagcc agacattaac gcttctggag aaactcaacg agctggacgc ggatgaacag
      541 gcagacatct gtgaatcgct tcacgaccac gctgatgagc tttaccgcag ctgcctcgcg
      601 cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct
      661 tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc
      721 gggtgtcggg gcgcagccat gacccagtca cgtagcgata gcggagtgta tactggctta
      781 actatgcggc atcagagcag attgtactga gagtgcacca ttgcggtgtg aaataccgca
      841 cagatgcgta aggagaaaat accgcatcag gcgctcttcc gcttcctcgc tcactgactc
      901 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
      961 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     1021 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     1081 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     1141 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     1201 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     1261 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     1321 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     1381 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     1441 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac
     1501 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     1561 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     1621 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     1681 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     1741 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     1801 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     1861 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     1921 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     1981 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     2041 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     2101 taatagtttg cgcaacgttg ttgccattgc tgcaggcatc gtggtgtcac gctcgtcgtt
     2161 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     2221 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     2281 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     2341 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     2401 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     2461 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     2521 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     2581 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     2641 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     2701 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     2761 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctaaat tgtaagcgtt
     2821 aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag
     2881 gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt
     2941 gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga
     3001 aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg
     3061 gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct
     3121 tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc
     3181 gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt
     3241 aatgcgccgc tacagggcgc gtcccattcg ccaatccgga tatagttcct cctttcagca
     3301 aaaaacccct caagacccgt ttagaggccc caaggggtta tgctagttat tgctcagcgg
     3361 tggcagcagc caactcagct tcctttcggg ctttgttagc agccggatct cagtggtggt
     3421 ggtggtggtg ctcgagtcat ttgtacagtt catccatacc atgcgtgatg cccgctgcgg
     3481 ttacgaactc cagcagaacc atatgatcgc gtttctcgtt cggatcttta gacagaacgc
     3541 tttgcgtgct cagatagtga ttgtctggca gcagaacagg accatcaccg attggagtgt
     3601 tttgctggta gtgatcagcc agctgcacgc tgccatcctc cacgttgtgg cgaattttaa
     3661 aattcgcttt aatgccattt ttttgtttat cggcggtgat gtaaacattg tggctgttaa
     3721 aattgtattc cagcttatgg cccaggatat tgccgtcttc tttaaagtca atgcctttca
     3781 gctcaatgcg gtttaccagg gtatcgcctt caaatttcac ttccgcacgc gttttgtacg
     3841 tgccgtcatc cttaaaggaa atcgtgcgtt cctgcacata gccttccggc atggcggact
     3901 tgaagaagtc atgctgcttc atatggtccg gataacgagc aaagcactga acaccataag
     3961 tcagcgtcgt taccagagtc ggccaaggta ccggcagttt accagtagta cagatgaact
     4021 tcagcgtcag tttaccatta gttgcgtcac cttcaccctc gccacgcacg gaaaacttat
     4081 gaccgttgac atcaccatcc agttccacca gaatagggac gacaccagtg aacagctctt
     4141 cgcctttacg catctagtat ttctcctctt taatctctag cttaaagtta attttgactg
     4201 gctgtgataa aggttaaacc ttccagcaag gggaaggtca agaaattaat aaaccaggcg
     4261 ggtaaaagtc cgtaaagatt aaaaaatcgg ctcgatttgc atcaggatta gacatttatc
     4321 tctttgtttt cctgtagtta agttgcgggt gctaagttaa atcaggatgc ctgaaaatcg
     4381 gcaccggggt gaggaattac ctcccgcatc tataaaaagg agttaacaaa agacgcgtca
     4441 gtgggctgat cattaactat ccgctggatg accaggatgc cattgctgtg gaagctgcct
     4501 gcactaatgt tccggcgtta tttcttgatg tctctgacca gacacccatc aacagtatta
     4561 ttttctccca tgaagacggt acgcgactgg gcgtggagca tctggtcgca ttgggtcacc
     4621 agcaaatcgc gctgttagcg ggcccattaa gttctgtctc ggcgcgtctg cgtctggctg
     4681 gctggcataa atatctcact cgcaatcaaa ttcagccgat agcggaacgg gaaggcgact
     4741 ggagtgccat gtccggtttt caacaaacca tgcaaatgct gaatgagggc atcgttccca
     4801 ctgcgatgct ggttgccaac gatcagatgg cgctgggcgc aatgcgcgcc attaccgagt
     4861 ccgggctgcg cgttggtgcg gatatctcgg tagtgggata cgacgatacc gaagacagct
     4921 catgttatat cccgccgtta accaccatca aacaggattt tcgcctgctg gggcaaacca
     4981 gcgtggaccg cttgctgcaa ctctctcagg gccaggcggt gaagggcaat cagctgttgc
     5041 ccgtctcact ggtgaaaaga aaaaccaccc tggcgcccaa tacgcaaacc gcctctcccc
     5101 gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc
     5161 agtgagcgca acgcaattaa tgtaagttag ctcactcatt aggcaccggg atctcgaccg
     5221 atgcccttga gagccttcaa cccagtcagc tccttccggt gggcgcgggg catgactatc
     5281 gtcgccgcac ttatgactgt cttctttatc atgcaactcg taggacaggt gccggcagcg
     5341 ctctgggtca ttttcggcga ggaccgcttt cgctggagcg cgacgatgat cggcctgtcg
     5401 cttgcggtat tcggaatctt gcacgccctc gctcaagcct tcgtcactgg tcccgccacc
     5461 aaacgtttcg gcgagaagca ggccattatc gccggcatgg cggccccacg ggtgcgcatg
     5521 atcgtgctcc tgtcg
//
