LOCUS       sequence-460527-        5514 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 5514)
  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 5514)
  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 mscarlet
FEATURES             Location/Qualifiers
     source          1..5514
                     /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"
     RBS             4139..4150
                     /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 ctcgagttat ttgtatagtt catccatgcc accggtagaa tgacgaccct
     3481 cggagcgctc atattgctct accaccgtat aatcttcatt atgagaggtg atgtccagtt
     3541 tgcggtccac attgtacgcg ccaggcatct gcacaggttt cttggctttg tacgtggttt
     3601 tgaagtcagc cagatagcgg ccgccatctt tcaggcgcag ggccatttta atgtcgcctt
     3661 tcagcacacc atcttccgga tacaagcgtt cggtagacgc ctcccaaccc atcgtctttt
     3721 tctgcatcac cgggccatca ggtggaaaat tagtaccacg cagtttaact ttatagatca
     3781 gggtgccatc ctccagggag gtgtcctgag tgacggtcac ggcaccacca tcttcaaaat
     3841 tcattacgcg ttcccacttg aaaccttccg gaaaagactg cttataatag tccgggatat
     3901 ccgctgggtg tttgatgaac gccctagaac cgtacataaa ttgcgggctc aggatgtccc
     3961 aggaaaaagg cagcgggccg cctttagtca ctttcagttt ggcggtctgg gtgccttcat
     4021 acggacggcc ctcgccttcg ccttcgatct cgaactcgtg accgttcata gaaccctcca
     4081 tgtgaacttt gaagcgcatg aactctttaa tcacagcttc tcctttactc atctagtatt
     4141 tctcctcttt aatctctagc ttaaagttaa ttttgactgg ctgtgataaa ggttaaacct
     4201 tccagcaagg ggaaggtcaa gaaattaata aaccaggcgg gtaaaagtcc gtaaagatta
     4261 aaaaatcggc tcgatttgca tcaggattag acatttatct ctttgttttc ctgtagttaa
     4321 gttgcgggtg ctaagttaaa tcaggatgcc tgaaaatcgg caccggggtg aggaattacc
     4381 tcccgcatct ataaaaagga gttaacaaaa gacgcgtcag tgggctgatc attaactatc
     4441 cgctggatga ccaggatgcc attgctgtgg aagctgcctg cactaatgtt ccggcgttat
     4501 ttcttgatgt ctctgaccag acacccatca acagtattat tttctcccat gaagacggta
     4561 cgcgactggg cgtggagcat ctggtcgcat tgggtcacca gcaaatcgcg ctgttagcgg
     4621 gcccattaag ttctgtctcg gcgcgtctgc gtctggctgg ctggcataaa tatctcactc
     4681 gcaatcaaat tcagccgata gcggaacggg aaggcgactg gagtgccatg tccggttttc
     4741 aacaaaccat gcaaatgctg aatgagggca tcgttcccac tgcgatgctg gttgccaacg
     4801 atcagatggc gctgggcgca atgcgcgcca ttaccgagtc cgggctgcgc gttggtgcgg
     4861 atatctcggt agtgggatac gacgataccg aagacagctc atgttatatc ccgccgttaa
     4921 ccaccatcaa acaggatttt cgcctgctgg ggcaaaccag cgtggaccgc ttgctgcaac
     4981 tctctcaggg ccaggcggtg aagggcaatc agctgttgcc cgtctcactg gtgaaaagaa
     5041 aaaccaccct ggcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa
     5101 tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat
     5161 gtaagttagc tcactcatta ggcaccggga tctcgaccga tgcccttgag agccttcaac
     5221 ccagtcagct ccttccggtg ggcgcggggc atgactatcg tcgccgcact tatgactgtc
     5281 ttctttatca tgcaactcgt aggacaggtg ccggcagcgc tctgggtcat tttcggcgag
     5341 gaccgctttc gctggagcgc gacgatgatc ggcctgtcgc ttgcggtatt cggaatcttg
     5401 cacgccctcg ctcaagcctt cgtcactggt cccgccacca aacgtttcgg cgagaagcag
     5461 gccattatcg ccggcatggc ggccccacgg gtgcgcatga tcgtgctcct gtcg
//
