LOCUS       sequence-50645-e        8200 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8200)
  AUTHORS   Filonov GS, Verkhusha VV
  TITLE     A Near-Infrared BiFC Reporter for In Vivo Imaging of Protein-Protein
            Interactions.
  JOURNAL   Chem Biol. 2013 Jul 23. pii: S1074-5521(13)00242-1. doi: 
            10.1016/j.chembiol.2013.06.009.
  PUBMED    23891149
REFERENCE   2  (bases 1 to 8200)
  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: "Chem Biol. 
            2013 Jul 23. pii: S1074-5521(13)00242-1. doi: 
            10.1016/j.chembiol.2013.06.009."
COMMENT     Sequence Label: pWA PAS-E BAD K-GAFm
FEATURES             Location/Qualifiers
     source          1..8200
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      822..908
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1018..1045
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        1064..1155
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1156..1224,1225..2016)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(1374..1393)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2187..2775
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2676..2695
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2929..2946
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2961..3103
                     /label=bom
                     /note="basis of mobility region from pBR322"
     CDS             complement(3205..3396)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     5038..5057
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5038..5056
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        5068..5186
                     /label=rhaB promoter
                     /note="promoter of the E. coli rhaBAD operon, conferring 
                     tight induction with L-rhamnose and repression with 
                     D-glucose in the presence of RhaR and RhaS (Giacalone et 
                     al., 2006)"
     CDS             5258..5275
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      6392..6478
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(6851..7729)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        7756..8040
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     7963..7982
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
ORIGIN
        1 cctccccagc gggacgtcgc cgagccgcag gcgttcttcc gccgcaccaa cagcgccatc
       61 cgccgcctgc aggccgccga aaccttggaa agcgcctgcg ccgccgcggc gcaagaggtg
      121 cggaagatta ccggctacga tcgggtgatg atctatcgct tcgcctccga cttcagcggc
      181 gaagtgatcg cagaggatcg gtgcgccgag gtcgagtcaa aactaggcct gcactatcct
      241 gcctcaaccg tgccggcgca ggcccgtcgg ctctatacca tcaacccggt acggatcatt
      301 cccgatatca attatcggcc ggtgccggtc accccatacc tcaatccggt caccgggcgg
      361 ccgattgatc ttagcttcgc catcctgcgc agcgtctcgc ccgtccatct ggaattcatg
      421 cgcaacatag gcatgcacgg cacgatgtcg atctcgattt tgcgcggcga gcgactgtgg
      481 ggattgatcg tttgccatca ccgaacgccg tactacgtcg atctcgatgg ccgccaagcc
      541 tgcgagctag tcgcccaggt tctggcccgg cagatcggcg tgatggaaga gtaaggtacc
      601 atatgggaat tcgaagcttg gctgttttgg cggatgagag aagattttca gcctgataca
      661 gattaaatca gaacgcagaa gcggtctgat aaaacagaat ttgcctggcg gcagtagcgc
      721 ggtggtccca cctgacccca tgccgaactc agaagtgaaa cgccgtagcg ccgatggtag
      781 tgtggggtct ccccatgcga gagtagggaa ctgccaggca tcaaataaaa cgaaaggctc
      841 agtcgaaaga ctgggccttt cgttttatct gttgtttgtc ggtgaacgct ctcctgagta
      901 ggacaaatcc gccgggagcg gatttgaacg ttgcgaagca acggcccgga gggtggcggc
      961 ggccgcgagg gtggcgggca ggacgcccgc cataaactgc caggcatcaa attaagcaga
     1021 aggccatcct gacggatggc ctttttgcgt ttctacaaac tcttttgttt atttttctaa
     1081 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
     1141 tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
     1201 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
     1261 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
     1321 gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt
     1381 ggcgcggtat tatcccgtgt tgacgccggg caagagcaac tcggtcgccg catacactat
     1441 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg
     1501 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
     1561 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
     1621 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
     1681 cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa
     1741 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca
     1801 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
     1861 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
     1921 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
     1981 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat
     2041 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     2101 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     2161 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     2221 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     2281 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta
     2341 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     2401 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     2461 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     2521 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     2581 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     2641 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     2701 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     2761 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     2821 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
     2881 gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg
     2941 agcgaggaag cggaagagcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt
     3001 tcacaccgca tatatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc
     3061 agtatacact ccgctatcgc tacgtgactg ggtcatggct gcgccccgac acccgccaac
     3121 acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt
     3181 gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag
     3241 gcagctgcgg taaagctcat cagcgtggtc gtgaagcgat tcacagatgt ctgcctgttc
     3301 atccgcgtcc agctcgttga gtttctccag aagcgttaat gtctggcttc tgataaagcg
     3361 ggccatgtta agggcggttt tttcctgttt ggtcacttga tgcctccgtg taagggggaa
     3421 tttctgttca tgggggtaat gataccgatg aaacgagaga ggatgctcac gatacgggtt
     3481 actgatgatg aacatgcccg gttactggaa cgttgtgagg gtaaacaact ggcggtatgg
     3541 atgcggcggg accagagaaa aatcactcag ggtcaatgcc agcgcttcgt taatacagat
     3601 gtaggtgttc cacagggtag ccagcagcat cctgcgatgc agatccggaa cataatggtg
     3661 cagggcgctg acttccgcgt ttccagactt tacgaaacac ggaaaccgaa gaccattcat
     3721 gttgttgctc aggtcgcaga cgttttgcag cagcagtcgc ttcacgttcg ctcgcgtatc
     3781 ggtgattcat tctgctaacc agtaaggcaa ccccgccagc ctagccgggt cctcaacgac
     3841 aggagcacga tcatgcgcac ccgtggccag gacccaacgc tgcccgagat gcgccgcgtg
     3901 cggctgctgg agatggcgga cgcgatggat atgttctgcc aagggttggt ttgcgcattc
     3961 acagttctcc gcaagaattg attggctcca attcttggag tggtgaatcc gttagcgagg
     4021 tgccgccggc ttccattcag gtcgaggtgg cccggctcca tgcaccgcga cgcaacgcgg
     4081 ggaggcagac aaggtatagg gcggcgccta caatccatgc caacccgttc catgtgctcg
     4141 ccgaggcggc ataaatcgcc gtgacgatca gcggtccagt gatcgaagtt aggctggtaa
     4201 gagccgcgag cgatccttga agctgtccct gatggtcgtc atctacctgc ctggacagca
     4261 tggcctgcaa cgcgggcatc ccgatgccgc cggaagcgag aagaatcata atggggaagg
     4321 ccatccagcc tcgcgtcgcg aacgccagca agacgtagcc cagcgcgtcg gccgccatgc
     4381 cggcgataat ggcctgcttc tcgccgaaac gtttggtggc gggaccagtg acgaaggctt
     4441 gagcgagggc gtgcaagatt ccgaataccg caagcgacag gccgatcatc gtcgcgctcc
     4501 agcgaaagcg gtcctcgccg aaaatgaccc agagcgctgc cggcacctgt cctacgagtt
     4561 gcatgataaa gaagacagtc ataagtgcgg cgacgatagt catgccccgc gcccaccgga
     4621 aggagctgac tgggttgaag gctctcaagg gcatcggtcg acgctctccc ttatgcgact
     4681 cctgcattag gaagcagccc agtagtaggt tgaggccgtt gagcaccgcc gccgcaagga
     4741 atggtgcatg ctcgatggct acgagggcag acagtaagtg gatttaccat aatcccttaa
     4801 ttgtacgcac cgctaaaacg cgttcagcgc gatcacggca gcagacaggt aaaaatggca
     4861 acaaaccacc ctaaaaactg cgcgatcgcg cctgataaat tttaaccgta tgaataccta
     4921 tgcaaccaga gggtacaggc cacattaccc ccacttaatc cactgaagct gccatttttc
     4981 atggtttcac catcccagcg aagggccatg catgcatcga aattaatacg acgaaattaa
     5041 tacgactcac tatagggcaa ttgcgatcac cacaattcag caaattgtga acatcatcac
     5101 gttcatcttt ccctggttgc caatggccca ttttcctgtc agtaacgaga aggtcgcgaa
     5161 ttcaggcgct ttttagactg gtcgtaggga gaccacaacg gtttccctct agaaataatt
     5221 ttcttaagaa ggagatatac atatgaccat gattacgcat catcatcatc atcatggctg
     5281 cagcatggcg gaaggatccg tcgccaggca gcctgacctc ttgacctgcg acgatgagcc
     5341 gatccatatc cccggtgcca tccaaccgca tggactgctg ctcgccctcg ccgccgacat
     5401 gacgatcgtt gccggcagcg acaaccttcc cgaactcacc ggactggcga tcggcgccct
     5461 gatcggccgc tctgcggccg atgtcttcga ctcggagacg cacaaccgtc tgacgatcgc
     5521 cttggccgag cccggggcgg ccgtcggagc accgatcact gtcggcttca cgatgcgaaa
     5581 ggacgcaggc ttcatcggct cctggcatcg ccatgatcag ctcatcttcc tcgagctcga
     5641 gcctccccag cggggcggca gcgaagtgag cgcgctggaa aaagaggtct ccgctcttga
     5701 gaaggaagtt tctgccttgg agaaagaggt gagcgccctg gagaaggaag tctctgcgct
     5761 tgaaaaaggc ggcagctaaa gatctcaggt tcaatggaag cgattaacga gtcgctccat
     5821 ccaccgatgg ataatcgggt acgcgaggct tgtcagtaca tcagcgatca cctggcagac
     5881 agcaattttg atatcgccag cgtcgcacag catgtttgct tgtcgccgtc gcgtctgtca
     5941 catcttttcc gccagcagtt agggattagc gtcttaagct ggcgcgagga ccaacgtatc
     6001 agccaggcga agctgctttt gagcaccacc cggatgccta tcgccaccgt cggtcgcaat
     6061 gttggttttg acgatcaact ctatttctcg cgggtattta aaaaatgcac cggggccagc
     6121 ccgagcgagt tccgtgccgg ttgtgaagaa aaagtgaatg atgtagccgt caagttgtca
     6181 taaaagcttg gctgttttgg cggatgagag aagattttca gcctgataca gattaaatca
     6241 gaacgcagaa gcggtctgat aaaacagaat ttgcctggcg gcagtagcgc ggtggtccca
     6301 cctgacccca tgccgaactc agaagtgaaa cgccgtagcg ccgatggtag tgtggggtct
     6361 ccccatgcga gagtagggaa ctgccaggca tcaaataaaa cgaaaggctc agtcgaaaga
     6421 ctgggccttt cgttttatct gttgtttgtc ggtgaacgct ctcctgagta ggacaaatcc
     6481 gccgggagcg gatttgacct aggcactctc agtacaatct gctctgatgc cgcatagtta
     6541 agccagtata cactccgcta tcgctacgtg actgggtcat ggctgcgccc cgacacccgc
     6601 caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag
     6661 ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg
     6721 cgaggcagca gatcaattcg cgcgcgaagg cgaagcggca tgcataatgt gcctgtcaaa
     6781 tggacgaagc agggattctg caaaccctat gctactccgt caagccgtca attgtctgat
     6841 tcgttaccaa ttatgacaac ttgacggcta catcattcac tttttcttca caaccggcac
     6901 ggaactcgct cgggctggcc ccggtgcatt ttttaaatac ccgcgagaaa tagagttgat
     6961 cgtcaaaacc aacattgcga ccgacggtgg cgataggcat ccgggtggtg ctcaaaagca
     7021 gcttcgcctg gctgatacgt tggtcctcgc gccagcttaa gacgctaatc cctaactgct
     7081 ggcggaaaag atgtgacaga cgcgacggcg acaagcaaac atgctgtgcg acgctggcga
     7141 tatcaaaatt gctgtctgcc aggtgatcgc tgatgtactg acaagcctcg cgtacccgat
     7201 tatccatcgg tggatggagc gactcgttaa tcgcttccat gcgccgcagt aacaattgct
     7261 caagcagatt tatcgccagc agctccgaat agcgcccttc cccttgcccg gcgttaatga
     7321 tttgcccaaa caggtcgctg aaatgcggct ggtgcgcttc atccgggcga aagaaccccg
     7381 tattggcaaa tattgacggc cagttaagcc attcatgcca gtaggcgcgc ggacgaaagt
     7441 aaacccactg gtgataccat tcgcgagcct ccggatgacg accgtagtga tgaatctctc
     7501 ctggcgggaa cagcaaaata tcacccggtc ggcaaacaaa ttctcgtccc tgatttttca
     7561 ccaccccctg accgcgaatg gtgagattga gaatataacc tttcattccc agcggtcggt
     7621 cgataaaaaa atcgagataa ccgttggcct caatcggcgt taaacccgcc accagatggg
     7681 cattaaacga gtatcccggc agcaggggat cattttgcgc ttcagccata cttttcatac
     7741 tcccgccatt cagagaagaa accaattgtc catattgcat cagacattgc cgtcactgcg
     7801 tcttttactg gctcttctcg ctaaccaaac cggtaacccc gcttattaaa agcattctgt
     7861 aacaaagcgg gaccaaagcc atgacaaaaa cgcgtaacaa aagtgtctat aatcacggca
     7921 gaaaagtcca cattgattat ttgcacggcg tcacactttg ctatgccata gcatttttat
     7981 ccataagatt agcggatcct acctgacgct ttttatcgca actctctact gtttctccat
     8041 acccgttttt ttgggctaac aggaggaatt aaccatgggc ggcagcaagg tgtctgcact
     8101 taaagaaaag gtttccgcac tcaaggagaa agtttcagct ttgaaagaga aggtcagtgc
     8161 ccttaaggaa aaagtgagcg cgctgaaaga aggcggcagc
//
