LOCUS       sequence-491598-        7872 bp    DNA     circular SYN 04-NOV-2025
DEFINITION  Re-NF-FF-Controller in high-copy backbone.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7872)
  TITLE     Re-NF-FF-Controller
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7872)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Nov 4, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: circuits C53-H
FEATURES             Location/Qualifiers
     source          1..7872
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             832..843
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             complement(855..1532)
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPYDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     RBS             1539..1550
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     terminator      1899..1970
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1986..2013
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     promoter        2082..2116
                     /label=BBa_J23106
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.47"
     RBS             2125..2136
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             2143..2856
                     /codon_start=1
                     /product="S65T variant of Aequorea victoria green 
                     fluorescent protein (Heim et al., 1995)"
                     /label=GFP (S65T)
                     /note="excitable with blue light"
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(2173..2201)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     2796..2816
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      2879..2950
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2966..2993
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     promoter        3008..3042
                     /label=BBa_J23110
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.33"
     RBS             3051..3062
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             3069..3947
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     terminator      3969..4040
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      4056..4083
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     promoter        4098..4132
                     /label=BBa_J23105
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.24"
     CDS             4161..5663
                     /codon_start=1
                     /transl_table=11
                     /gene="Bxb1"
                     /product="Bxb1 integrase"
                     /label=Bxb1 integrase
                     /note="Bxb1 large serine integrase from Mycobacterium 
                     bacteriophage Bxb1 (Ghosh et al. 2003)."
                     /protein_id="AFF61396.1"
                     /translation="MRALVVIRLSRVTDATTSPERQLEYCQQLCAQRGWDVVGVAEDLD
                     VSGAVDPFDRKRRPNLARWLAFEEQPFDVIVAYRVDRLTRSIRHLQQLVHWAEDHKKLV
                     VSATEAHFDTTTPFAAVVIALMGTVAQMELEAIKERNRSAAHFNIRAGKYRGSLPPWGY
                     LPTRVDGEWRLVPDPVQRERILEVYHRVVDNHEPLHLVAHDLNRRGVLSPKDYFAQLQG
                     REPQGREWSATALKRSMISEAMLGYATLNGKTVRDDDGAPLVRAEPILTREQLEALRAE
                     LVKTSRAKPAVSTPSLLLRVLFCAVCGEPAYKFAGGGRKHPRYRCRSMGFPKHCGNGTV
                     AMAEWDAFCEEQVLDLLGDAERLEKVWVAGSDSAVELAEVNAELVDLTSLVGSPAYRAG
                     SPQREALDARIAALAARQEELEGLEARPSGWEWRETGQRFGDWWREQDTAAKNTWLRSM
                     NVRLTFDVRGGLTRTIDFGDLQEYEQHLRLGSVVERLHTGMS"
     terminator      5682..5753
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      5769..5796
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     misc_feature    5803..5823
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      5824..5881
                     /label=his operon terminator
                     /note="This putative transcriptin terminator from the E. 
                     coli his operon has a 2-bp deletion introduced during 
                     synthesis. Its efficiency has not been determined."
     primer_bind     complement(5905..5922)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6076..6664)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6156..6175)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(6847..6941)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(6962..7621)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     7536..7555
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     primer_bind     7783..7801
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(7805..7848)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    7851..7872
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
ORIGIN
        1 gcgaaaaaac cccgccgaag cggggttttt tgcgtactag agttattaac cggtgacttc
       61 ccagacctgg ccgtcaacgg tgaacttgcc tcccaggatc gggacgatct cagccttgac
      121 atgcttgccg tcgaccgtga gcatcccgaa gcccatctgc cagttcccgg ctccgccctt
      181 tagatagttg gccttcttca tgtccatcag gtgcccgact tccatgccgg tgacggtctt
      241 gcgcaccgag ccgccgtacc cgaacgagtg cgagacgaca gcctgccggt gcgtgtggcc
      301 gcagaccacg gacttgccga acttcttggc accgttgagc gctgtcgatc cggcgatctg
      361 ggatagcgtc atcttgccca tgtgcccgtg agtggagatc cagcccggag cgatgtcgta
      421 gaagtcaggc agcagctcca caccgaaccc gtcgaagtcg agcagcacgt cgatgtcgaa
      481 agcgtgcgta ccctccaggg ccggtgcgtt cttggccagg tactcgcggg ctcgcagatc
      541 gtggttgccc tcgtgcatcc cgatccagcc gtcgtagacc ttgcgcagcg gctccatcag
      601 gttcttcttg gcgtagtccg cgtcgcggta caccgagccc tcgaactcgc ccttggttcc
      661 ccggttccag cgcgagggct gcggcaggtc taggacgtca ccgatatgta ccacgccgta
      721 cggctggaca tccccgatga agcggatgac cgcttgcatc tctttgcgcg cctcgaaagg
      781 caactgagta tcgggtagaa agacgatacg ctgagtgccg gccatctagt atttctcctc
      841 ttttactaga gttattaagc accggtggag tgacgacctt cagcacgttc gtactgttca
      901 acgatggtgt agtcttcgtt gtgggaggtg atgtccagtt tgatgtcggt tttgtaagca
      961 cccggcagct gaaccggttt tttagccatg taggtggttt taacttcagc gtcgtagtga
     1021 ccaccgtctt tcagtttcag acgcattttg atttcacctt tcagagcacc gtcttccggg
     1081 tacatacgtt cggtggaagc ttcccaaccc atggtttttt tctgcataac cggaccgtcg
     1141 tacgggaagt tggtaccacg cagtttaact ttgtagatga actcaccgtc ttgcagggag
     1201 gagtcctggg taacggtaac aacaccaccg tcttcgaagt tcataacacg ttcccatttg
     1261 aaaccttccg ggaaggacag tttcaggtag tccgggatgt cagccgggtg tttaacgtaa
     1321 gctttggaac cgtactggaa ctgcggggac aggatgtccc aagcgaacgg cagcggacca
     1381 cctttggtaa ctttcagttt agcggtctgg gtaccttcgt acggacgacc ttcaccttca
     1441 ccttcgattt cgaactcgtg accgttaacg gaaccttcca tacgaacttt gaaacgcatg
     1501 aactctttga taacgtcttc ggaggaagcc atctagtatt tctcctcttt tactagagcg
     1561 gccggcttgt cgacgacggc gcactcagtg gtgtacggta caaaccccga caatagcgag
     1621 gaggtaaatt gtaagcttta gggaaacgac atttactgac gcgactttct caaagctagg
     1681 gtgttggcct gaactgtacg cttggagttg taggacagac ataaaagtat cagatacgca
     1741 cactaaccat ctgctagccc aaaaaaacgg tatggagaaa cagtagagag ttgcgataaa
     1801 aagcgtcagg taggatccgc taatcttatg gactatcttg ctatggcata gcaaagtgtg
     1861 acgccgtgca aataatcaat gtctctagta ccaggcatca aataaaacga aaggctcagt
     1921 cgaaagactg ggcctttcgt tttatctgtt gtttgtcggt gaacgctctc tactagagtc
     1981 acactggctc accttcgggt gggcctttct gcgtttatag cccggatgat cctgacgacg
     2041 gagtgcgcgg tggttgacca gacaaaccac gactactaga gtttacggct agctcagtcc
     2101 taggtatagt gctagctact agagaaagag gagaaatact agatgcgtaa aggagaagaa
     2161 cttttcactg gagttgtccc aattcttgtt gaattagatg gtgatgttaa tgggcacaaa
     2221 ttttctgtca gtggagaggg tgaaggtgat gcaacatacg gaaaacttac ccttaaattt
     2281 atttgcacta ctggaaaact acctgttcca tggccaacac ttgtcactac tttcggttat
     2341 ggtgttcaat gctttgcgag atacccagat catatgaaac agcatgactt tttcaagagt
     2401 gccatgcccg aaggttatgt acaggaaaga actatatttt tcaaagatga cgggaactac
     2461 aagacacgtg ctgaagtcaa gtttgaaggt gatacccttg ttaatagaat cgagttaaaa
     2521 ggtattgatt ttaaagaaga tggaaacatt cttggacaca aattggaata caactataac
     2581 tcacacaatg tatacatcat ggcagacaaa caaaagaatg gaatcaaagt taacttcaaa
     2641 attagacaca acattgaaga tggaagcgtt caactagcag accattatca acaaaatact
     2701 ccaattggcg atggccctgt ccttttacca gacaaccatt acctgtccac acaatctgcc
     2761 ctttcgaaag atcccaacga aaagagagac cacatggtcc ttcttgagtt tgtaacagct
     2821 gctgggatta cacatggcat ggatgaacta tacaaataat aatactagag ccaggcatca
     2881 aataaaacga aaggctcagt cgaaagactg ggcctttcgt tttatctgtt gtttgtcggt
     2941 gaacgctctc tactagagtc acactggctc accttcgggt gggcctttct gcgtttatat
     3001 actagagttt acggctagct cagtcctagg tacaatgcta gctactagag aaagaggaga
     3061 aatactagat ggctgaagcg caaaatgatc ccctgctgcc gggatactcg tttaacgccc
     3121 atctggtggc gggtttaacg ccgattgagg ccaatggtta tctcgatttt tttatcgacc
     3181 gaccgctggg aatgaaaggt tatattctca atctcaccat tcgcggtcag ggggtggtga
     3241 aaaatcaggg acgagaattt gtctgccgac cgggtgatat tttgctgttc ccgccaggag
     3301 agattcatca ctacggtcgt catccggagg ctcgcgaatg gtatcaccag tgggtttact
     3361 ttcgtccgcg cgcctactgg catgaatggc ttaactggcc gtcaatattt gccaatacgg
     3421 gtttctttcg cccggatgaa gcgcaccagc cgcatttcag cgacctgttt gggcaaatca
     3481 ttaacgccgg gcaaggggaa gggcgctatt cggagctgct ggcgataaat ctgcttgagc
     3541 aattgttact gcggcgcatg gaagcgatta acgagtcgct ccatccaccg atggataatc
     3601 gggtacgcga ggcttgtcag tacatcagcg atcacctggc agacagcaat tttgatatcg
     3661 ccagcgtcgc acagcatgtt tgcctgtcgc cgtcgcgtct gtcacatctt ttccgccagc
     3721 agttagggat tagcgtctta agctggcgcg aggaccaacg catcagccag gcgaagctgc
     3781 ttttgagcac tacccggatg cctatcgcca ccgtcggtcg caatgttggt tttgacgatc
     3841 aactctattt ctcgcgagta tttaaaaaat gcaccggggc cagcccgagc gagttccgtg
     3901 ccggttgtga agaaaaagtg aatgatgtag ccgtcaagtt gtcataataa actactagag
     3961 ccaggcatca aataaaacga aaggctcagt cgaaagactg ggcctttcgt tttatctgtt
     4021 gtttgtcggt gaacgctctc tactagagtc acactggctc accttcgggt gggcctttct
     4081 gcgtttatat actagagttt acggctagct cagtcctagg tactatgcta gctactagag
     4141 tcacacagga aacctactag atgagagccc tggtagtcat ccgcctgtcc cgcgtcaccg
     4201 atgctacgac ttcaccggag cgtcagctgg agtattgcca gcagctctgc gcccagcgcg
     4261 gctgggacgt cgtcggggta gcggaggatc tggacgtctc cggggcggtc gatccgttcg
     4321 accggaagcg cagaccgaac ctggcccggt ggctagcgtt cgaggagcaa ccgtttgacg
     4381 tgatcgtggc gtaccgggta gatcggttga cccgatcgat ccggcatctt cagcagctgg
     4441 tccactgggc cgaggaccac aagaagctgg tcgtctccgc gaccgaagcg cacttcgata
     4501 cgacgacgcc gtttgcggcg gtcgtcatcg cgcttatggg aacggtggcg cagatggaat
     4561 tagaagcgat caaagagcgg aaccgttcgg ctgcgcattt caatatccgt gccgggaaat
     4621 accgaggatc cctgccgccg tggggatacc tgcctacgcg cgtggacggg gagtggcggc
     4681 tggtgccgga ccctgtgcag cgagagcgca tcctcgaggt gtatcaccgc gtcgtcgaca
     4741 accacgagcc gctgcatctg gtggcccacg acctgaaccg gcgtggtgtc ctgtcgccga
     4801 aggactactt cgcgcagctg caaggccgcg agccgcaggg ccgggagtgg tcggctaccg
     4861 cgctgaagcg atcgatgatc tccgaggcga tgctcgggta cgcgactctg aacggtaaga
     4921 ccgtccgaga cgacgacgga gccccgctgg tgcgggctga gccgatcctg acccgtgagc
     4981 agctggaggc gctgcgcgcc gagctcgtga agacctcccg ggcgaagccc gcggtgtcta
     5041 ccccgtcgct gctgctgcgg gtgttgttct gcgcggtgtg cggggagccc gcgtacaagt
     5101 tcgccggggg aggacgtaag cacccgcgct accgctgccg ctcgatgggg ttcccgaagc
     5161 actgcgggaa cggcacggtg gcgatggccg agtgggacgc gttctgcgag gagcaggtac
     5221 tggatctgct cggggacgcg gagcgtctgg agaaagtctg ggtagcgggc tcggactccg
     5281 cggtcgaact cgcggaggtg aacgcggagc tggtggacct gacgtcgctg gtcggctccc
     5341 cggcctaccg ggcgggctct ccgcagcgag aagcactgga tgcccgtatt gcggcgctgg
     5401 ccgcgcggca agaggagctg gagggcctgg aggctcgccc gtctggctgg gagtggcgcg
     5461 agaccgggca gcggttcggg gactggtggc gggagcagga caccgcggca aagaacacct
     5521 ggcttcggtc gatgaacgtt cggctgacgt tcgacgtccg cggcgggctg actcgcacga
     5581 tcgacttcgg ggatcttcag gagtacgagc agcatctcag gctcggcagc gtggtcgaac
     5641 ggctacacac cgggatgtcg taataaacta gagccaggca tcaaataaaa cgaaaggctc
     5701 agtcgaaaga ctgggccttt cgttttatct gttgtttgtc ggtgaacgct ctctactaga
     5761 gtcacactgg ctcaccttcg ggttggcctt tctgcgttta tatactagta gcggccgctg
     5821 cagtccggca aaaaagggca aggtgtcacc accctgccct ttttctttaa aaccgaaaag
     5881 attacttcgc gttatgcagg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     5941 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     6001 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     6061 cgcgttgctg gcgtttttcc acaggctccg cccccctgac gagcatcaca aaaatcgacg
     6121 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     6181 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     6241 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     6301 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     6361 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     6421 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     6481 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     6541 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     6601 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     6661 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     6721 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     6781 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagctcgag
     6841 gcttggattc tcaccaataa aaaacgcccg gcggcaaccg agcgttctga acaaatccag
     6901 atggagttct gaggtcatta ctggatctat caacaggagt ccaagcgagc tcgatatcaa
     6961 attacgcccc gccctgccac tcatcgcagt actgttgtaa ttcattaagc attctgccga
     7021 catggaagcc atcacaaacg gcatgatgaa cctgaatcgc cagcggcatc agcaccttgt
     7081 cgccttgcgt ataatatttg cccatggtga aaacgggggc gaagaagttg tccatattgg
     7141 ccacgtttaa atcaaaactg gtgaaactca cccagggatt ggctgagacg aaaaacatat
     7201 tctcaataaa ccctttaggg aaataggcca ggttttcacc gtaacacgcc acatcttgcg
     7261 aatatatgtg tagaaactgc cggaaatcgt cgtggtattc actccagagc gatgaaaacg
     7321 tttcagtttg ctcatggaaa acggtgtaac aagggtgaac actatcccat atcaccagct
     7381 caccgtcttt cattgccata cgaaattccg gatgagcatt catcaggcgg gcaagaatgt
     7441 gaataaaggc cggataaaac ttgtgcttat ttttctttac ggtctttaaa aaggccgtaa
     7501 tatccagctg aacggtctgg ttataggtac attgagcaac tgactgaaat gcctcaaaat
     7561 gttctttacg atgccattgg gatatatcaa cggtggtata tccagtgatt tttttctcca
     7621 ttttagcttc cttagctcct gaaaatctcg ataactcaaa aaatacgccc ggtagtgatc
     7681 ttatttcatt atggtgaaag ttggaacctc ttacgtgccc gatcaactcg agtgccacct
     7741 gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg
     7801 cagaatttca gataaaaaaa atccttagct ttcgctaagg atgatttctg gaattcgcgg
     7861 ccgcttctag ag
//
