LOCUS       sequence-369315-        4948 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  sfGFP under light light responsive PEL222 promoter and EL222 under 
            constitutively active BBa_J2305 promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4948)
  AUTHORS   Pirhanov A, Bridges CM, Goodwin RA, Guo YS, Furrer J, Shor LM, Gage 
            DJ, Cho YK
  TITLE     Optogenetics in Sinorhizobium meliloti Enables Spatial Control of 
            Exopolysaccharide Production and Biofilm Structure.
  JOURNAL   ACS Synth Biol. 2021 Feb 19;10(2):345-356. doi: 
            10.1021/acssynbio.0c00498. Epub 2021 Jan 19.
  PUBMED    33465305
REFERENCE   2  (bases 1 to 4948)
  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.0c00498"; journalName: "ACS Synth Biol"; date: 
            "2021-02-19- 19"; volume: "10"; issue: "2"; pages: "345-356"
COMMENT     Sequence Label: pAP05
FEATURES             Location/Qualifiers
     source          1..4948
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             161..874
                     /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"
     terminator      961..1055
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     RBS             1186..1197
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             1887..3077
                     /codon_start=1
                     /gene="tet"
                     /product="tetracycline efflux protein"
                     /label=TcR
                     /note="confers resistance to tetracycline"
                     /translation="MKSNNALIVILGTVTLDAVGIGLVMPVLPGLLRDIVHSDSIASHY
                     GVLLALYALMQFLCAPVLGALSDRFGRRPVLLASLLGATIDYAIMATTPVLWILYAGRI
                     VAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHAPF
                     LAAAVLNGLNLLLGCFLMQESHKGERRPMPLRAFNPVSSFRWARGMTIVAALMTVFFIM
                     QLVGQVPAALWVIFGEDRFRWSATMSGLSLAVFGILHALAQAFVTGPATKRFGEKQAII
                     AGMAADALGYVLLAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSL
                     AALTSLTSITGPLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSRATST"
     primer_bind     complement(2239..2258)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     oriT            3223..3331
                     /note="incP origin of transfer"
     CDS             complement(3342..4004)
                     /codon_start=1
                     /product="replication protein for the broad-host-range 
                     plasmid pBBR1 from Bordetella bronchiseptica"
                     /label=pBBR1 Rep
                     /translation="MATQSREIGIQAKNKPGHWVQTERKAHEAWAGLIARKPTAAMLLH
                     HLVAQMGHQNAVVVSQKTLSKLIGRSLRTVQYAVKDLVAERWISVVKLNGPGTVSAYVV
                     NDRVAWGQPRDQLRLSVFSAAVVVDHDDQDESLLGHGDLRRIPTLYPGEQQLPTGPGEE
                     PPSQPGIPGMEPDLPALTETEEWERRGQQRLPMPDEPCFLDDGEPLEPPTRVTLPRR"
     rep_origin      4005..4776
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"
     terminator      complement(join(4891..4948,1..29))
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 tctttcgact gagcctttcg ttttatttga tgcctttaat taaagcgaat aacaatttca
       61 cacagagccg gtagccttta gtccatgtta gcgaagaaaa tggtttgtta tagtcgaata
      121 aacacacaga attcattaag aaagaaggag atatggtacc atgcgtaaag gcgaagagct
      181 gttcactggt gtcgtcccta ttctggtgga actggatggt gatgtcaacg gtcataagtt
      241 ttccgtgcgt ggcgagggtg aaggtgacgc aactaatggt aaactgacgc tgaagttcat
      301 ctgtactact ggtaaactgc cggtaccttg gccgactctg gtaacgacgc tgacttatgg
      361 tgttcagtgc tttgctcgtt atccggacca tatgaagcag catgacttct tcaagtccgc
      421 catgccggaa ggctatgtgc aggaacgcac gatttccttt aaggatgacg gcacgtacaa
      481 aacgcgtgcg gaagtgaaat ttgaaggcga taccctggta aaccgcattg agctgaaagg
      541 cattgacttt aaagaagacg gcaatatcct gggccataag ctggaataca attttaacag
      601 ccacaatgtt tacatcaccg ccgataaaca aaaaaatggc attaaagcga attttaaaat
      661 tcgccacaac gtggaggatg gcagcgtgca gctggctgat cactaccagc aaaacactcc
      721 aatcggtgat ggtcctgttc tgctgccaga caatcactat ctgagcacgc aaagcgttct
      781 gtctaaagat ccgaacgaga aacgcgatca tatggttctg ctggagttcg taaccgcagc
      841 gggcatcacg catggtatgg atgaactgta caaatgaaca cgggatcctc tagagtcgac
      901 ctgcaggcat gcaagcttgc ggccgcgtcg tgactgggaa aaccctggcg actagtcttg
      961 gactcctgtt gatagatcca gtaatgacct cagaactcca tctggatttg ttcagaacgc
     1021 tcggttgccg ccgggcgttt tttattggtg agaatccagg ggtccccaat aattacgatt
     1081 tacgtattta aatttgacag cttatcatcg ataaactgta atgcggtagt ttatcaagga
     1141 ggctagctca gtcctaggta ctatgctagc cacacagaat tcattaaaga ggagaaaggt
     1201 accatgttgg atatgggaca agatcggccg atcgatggaa gtggggcacc cggggcagac
     1261 gacacacgcg ttgaggtgca accgccggcg cagtgggtcc tcgacctgat cgaggccagc
     1321 ccgatcgcat cggtcgtgtc cgatccgcgt ctcgccgaca atccgctgat cgccatcaac
     1381 caggccttca ccgacctgac cggctattcc gaagaagaat gcgtcggccg caattgccga
     1441 ttcctggcag gttccggcac cgagccgtgg ctgaccgaca agatccgcca aggcgtgcgc
     1501 gagcacaagc cggtgctggt cgagatcctg aactacaaga aggacggcac gccgttccgc
     1561 aatgccgtgc tcgttgcacc gatctacgat gacgacgacg agcttctcta tttcctcggc
     1621 agccaggtcg aagtcgacga cgaccagccc aacatgggca tggcgcgccg cgaacgcgcc
     1681 gcggaaatgc tcaagacgct gtcgccgcgc cagctcgagg ttacgacgct ggtggcatcg
     1741 ggcttgcgca acaaggaagt ggcggcccgg ctcggcctgt cggagaaaac cgtcaagatg
     1801 caccgcgggc tggtgatgga aaagctcaac ctgaagacca gcgccgatct ggtgcgcatt
     1861 gccgtcgaag ccggaatctg acttctatga aatctaacaa tgcgctcatc gtcatcctcg
     1921 gcaccgtcac cctggatgct gtaggcatag gcttggttat gccggtactg ccgggcctct
     1981 tgcgggatat cgtccattcc gacagcatcg ccagtcacta tggcgtgctg ctagcgctat
     2041 atgcgttgat gcaatttcta tgcgcacccg ttctcggagc actgtccgac cgctttggcc
     2101 gccgcccagt cctgctcgct tcgctacttg gagccactat cgactacgcg atcatggcga
     2161 ccacacccgt cctgtggatt ctctacgccg gacgcatcgt ggccggcatc accggcgcca
     2221 caggtgcggt tgctggcgcc tatatcgccg acatcaccga tggggaagat cgggctcgcc
     2281 acttcgggct catgagcgct tgtttcggcg tgggtatggt ggcaggcccc gtggccgggg
     2341 gactgttggg cgccatctcc ctgcacgcac cattccttgc ggcggcggtg ctcaacggcc
     2401 tcaacctact actgggctgc ttcctaatgc aggagtcgca taagggagag cgccgtccga
     2461 tgcccttgag agccttcaac ccagtcagct ccttccggtg ggcgcggggc atgaccattg
     2521 tggccgcact tatgactgtc ttctttatca tgcaactcgt aggacaggtg ccggcagcgc
     2581 tctgggtcat tttcggcgag gaccgctttc gctggagcgc gacgatgagc ggcctgtcgc
     2641 ttgcggtatt cggaatcttg cacgccctcg ctcaagcctt cgtcactggt cccgccacca
     2701 aacgtttcgg cgagaagcag gccattatcg ccggcatggc ggccgacgcg ctgggctacg
     2761 tcttgctggc gttcgcgacg cgaggctgga tggccttccc cattatgatt cttctcgctt
     2821 ccggcggtat cgggatgccc gcgttgcagg ccatgctgtc caggcaggta gatgacgacc
     2881 atcagggaca gcttcaagga tcgctcgcgg ctcttaccag cctaacttcg atcactggac
     2941 cgctgatcgt cacggcgatt tatgccgcct cggcgagcac atggaacggg ttggcatgga
     3001 ttgtaggcgc cgccctatac cttgtctgcc tccccgcgtt gcgtcgcggt gcatggagcc
     3061 gggccacctc gacctgagac aattgtcctt ttccgctgca taaccctgct tcggggtcat
     3121 tatagcgatt ttttcggtat atccatcctt tttcgcacga tatacaggat tttgccaaag
     3181 ggttcgtgta gactttcctt ggtgtatcca acggcgtcag ccgggcagga taggtgaagt
     3241 aggcccaccc gcgagcgggt gttccttctt cactgtccct tattcgcacc tggcggtgct
     3301 caacgggaat cctgctctgc gaggctggcc gtaggccggc cctaccggcg cggcagcgtt
     3361 acccgtgtcg gcggctccaa cggctcgcca tcgtccagaa aacacggctc atcgggcatc
     3421 ggcaggcgct gctgcccgcg ccgttcccat tcctccgttt cggtcaaggc tggcaggtct
     3481 ggttccatgc ccggaatgcc gggctggctg ggcggctcct cgccggggcc ggtcggtagt
     3541 tgctgctcgc ccggatacag ggtcgggatg cggcgcaggt cgccatgccc caacagcgat
     3601 tcgtcctggt cgtcgtgatc aaccaccacg gcggcactga acaccgacag gcgcaactgg
     3661 tcgcggggct ggccccacgc cacgcggtca ttgaccacgt aggccgacac ggtgccgggg
     3721 ccgttgagct tcacgacgga gatccagcgc tcggccacca agtccttgac tgcgtattgg
     3781 accgtccgca aagaacgtcc gatgagcttg gaaagtgtct tctggctgac caccacggcg
     3841 ttctggtggc ccatctgcgc cacgaggtga tgcagcagca ttgccgccgt gggtttcctc
     3901 gcaataagcc cggcccacgc ctcatgcgct ttgcgttccg tttgcaccca gtgaccgggc
     3961 ttgttcttgg cttgaatgcc gatttctctg gactgcgtgg ccatgcttat ctccatgcgg
     4021 taggggtgcc gcacggttgc ggcaccatgc gcaatcagct gcaacttttc ggcagcgcga
     4081 caacaattat gcgttgcgta aaagtggcag tcaattacag attttcttta acctacgcaa
     4141 tgagctattg cggggggtgc cgcaatgagc tgttgcgtac cccccttttt taagttgttg
     4201 atttttaagt ctttcgcatt tcgccctata tctagttctt tggtgcccaa agaagggcac
     4261 ccctgcgggg ttcccccacg ccttcggcgc ggctccccct ccggcaaaaa gtggcccctc
     4321 cggggcttgt tgatcgactg cgcggccttc ggccttgccc aaggtggcgc tgcccccttg
     4381 gaacccccgc actcgccgcc gtgaggctcg gggggcaggc gggcgggctt cgcccttcga
     4441 ctgcccccac tcgcataggc ttgggtcgtt ccaggcgcgt caaggccaag ccgctgcgcg
     4501 gtcgctgcgc gagccttgac ccgccttcca cttggtgtcc aaccggcaag cgaagcgcgc
     4561 aggccgcagg ccggaggctt ttccccagag aaaattaaaa aaattgatgg ggcaaggccg
     4621 caggccgcgc agttggagcc ggtgggtatg tggtcgaagg ctgggtagcc ggtgggcaat
     4681 ccctgtggtc aagctcgtgg gcaggcgcag cctgtccatc agcttgtcca gcagggttgt
     4741 ccacgggccg agcgaagcga gccagccggt ggccgctcgc ggccatcgtc cacatatcca
     4801 cgggctggca agggagcgca gcgaccgcgc agggcgaagc ccggagagca agcccgtagg
     4861 gggggcgcgc ccagctgtct agggcggcgg atttgtccta ctcaggagag cgttcaccga
     4921 caaacaacag ataaaacgaa aggcccag
//
