LOCUS       sequence-8742-ex        6225 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 6225)
  AUTHORS   Pfleger BF, Fawzi NJ, Keasling JD
  TITLE     Optimization of DsRed production in Escherichia coli: effect of 
            ribosome binding site sequestration on translation efficiency.
  JOURNAL   Biotechnol Bioeng. 2005 Dec 5. 92(5):553-8.
  PUBMED    16247774
REFERENCE   2  (bases 1 to 6225)
  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: "Biotechnol 
            Bioeng. 2005 Dec 5. 92(5):553-8."
COMMENT     Sequence Label: pBAD*RFPEC2
FEATURES             Location/Qualifiers
     source          1..6225
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             128..844
                     /codon_start=1
                     /product="GFP variant optimized for excitation by UV light"
                     /label=GFPuv
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFSYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKA
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(158..186)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     781..801
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(1076..1093)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1246..1332
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1424..1451
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        1470..1561
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1562..1630,1631..2422)
                     /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(1780..1799)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2464..2919
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2551..2570)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     rep_origin      3030..3618
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3519..3538
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3772..3789
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     3955..3977
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4116..4135
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             complement(4315..5193)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        5220..5504
                     /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     5427..5446
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             join(5549..6225,1)
                     /codon_start=1
                     /product="rapidly maturing tetrameric variant of DsRed 
                     fluorescent protein (Bevis and Glick, 2002)"
                     /label=DsRed-Express
                     /note="mammalian codon-optimized"
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGSFIYKVKFIGVNFPSDGPVMQKKTMGWEASTERLYPRDGVLKGEIHK
                     ALKLKDGGHYLVEFKSIYMAKKPVQLPGYYYVDSKLDITSHNEDYTIVEQYERAEGRHH
                     LFL"
     primer_bind     complement(5679..5697)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     complement(5729..5749)
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     primer_bind     5947..5966
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     6141..6164
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
ORIGIN
        1 gatctgatcg ttgcgggcgg ggcgagagtc tcgccccgcc cgcgaccgcg gtgaaaatac
       61 gagaatatta tttgtattga tctcctaggc ggggtaccgt attttggatg ataacgaggc
      121 gcaaaaaatg agtaaaggag aagaactttt cactggagtt gtcccaattc ttgttgaatt
      181 agatggtgat gttaatgggc acaaattttc tgtcagtgga gagggtgaag gtgatgcaac
      241 atacggaaaa cttaccctta aatttatttg cactactgga aaactacctg ttccatggcc
      301 aacacttgtc actactttct cttatggtgt tcaatgcttt tcccgttatc cggatcatat
      361 gaaacggcat gactttttca agagtgccat gcccgaaggt tatgtacagg aacgcactat
      421 atctttcaaa gatgacggga actacaagac gcgtgctgaa gtcaagtttg aaggtgatac
      481 ccttgttaat cgtatcgagt taaaaggtat tgattttaaa gaagatggaa acattctcgg
      541 acacaaactg gagtacaact ataactcaca caatgtatac atcacggcag acaaacaaaa
      601 gaatggaatc aaagctaact tcaaaattcg ccacaacatt gaagatggat ccgttcaact
      661 agcagaccat tatcaacaaa atactccaat tggcgatggc cctgtccttt taccagacaa
      721 ccattacctg tccacacaat ctgccctttc gaaagatccc aacgaaaagc gtgaccacat
      781 ggtccttctt gagtttgtaa ctgctgctgg gattacacat ggcatggatg agctctacaa
      841 ataagatgaa ttccagctga gcgccggtcg ctaccattac cagttggtct ggtgtcaaaa
      901 ataataataa ccgggcaggg gggatctgtt aagttctcgt ctggtagaaa aacgctgctg
      961 cgggtttttt tttgccttta gtaaattgaa ctgactttcg tcagttattc cttacccagc
     1021 aatgcctgca ggcatgcaag cttggctgtt ttggcggatg agagaagatt ttcagcctga
     1081 tacagattaa atcagaacgc agaagcggtc tgataaaaca gaatttgcct ggcggcagta
     1141 gcgcggtggt cccacctgac cccatgccga actcagaagt gaaacgccgt agcgccgatg
     1201 gtagtgtggg gtctccccat gcgagagtag ggaactgcca ggcatcaaat aaaacgaaag
     1261 gctcagtcga aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa cgctctcctg
     1321 agtaggacaa atccgccggg agcggatttg aacgttgcga agcaacggcc cggagggtgg
     1381 cgggcaggac gcccgccata aactgccagg catcaaatta agcagaaggc catcctgacg
     1441 gatggccttt ttgcgtttct acaaactctt ttgtttattt ttctaaatac attcaaatat
     1501 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
     1561 tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc
     1621 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc
     1681 acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc
     1741 cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc
     1801 ccgtgttgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt
     1861 ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt
     1921 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat
     1981 cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct
     2041 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat
     2101 gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc
     2161 ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg
     2221 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc
     2281 tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta
     2341 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc
     2401 ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga
     2461 tttacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
     2521 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
     2581 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
     2641 ttagtgcttt acggcacctc gaccccaaaa aacttgattt gggtgatggt tcacgtagtg
     2701 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
     2761 gtggactctt gttccaaact tgaacaacac tcaaccctat ctcgggctat tcttttgatt
     2821 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
     2881 ttaacgcgaa ttttaacaaa atattaacgt ttacaattta aaaggatcta ggtgaagatc
     2941 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     3001 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     3061 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     3121 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt
     3181 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     3241 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     3301 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     3361 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     3421 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     3481 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     3541 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     3601 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     3661 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     3721 accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca
     3781 gtgagcgagg aagcggaaga gcgcctgatg cggtattttc tccttacgca tctgtgcggt
     3841 atttcacacc gcatagggtc atggctgcgc cccgacaccc gccaacaccc gctgacgcgc
     3901 cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga
     3961 gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgaggcag caaggagatg
     4021 gcgcccaaca gtcccccggc cacggggcct gccaccatac ccacgccgaa acaagcgctc
     4081 atgagcccga agtggcgagc ccgatcttcc ccatcggtga tgtcggcgat ataggcgcca
     4141 gcaaccgcac ctgtggcgcc ggtgatgccg gccacgatgc gtccggcgta gaggatctgc
     4201 tcatgtttga cagcttatca tcgatgcata atgtgcctgt caaatggacg aagcagggat
     4261 tctgcaaacc ctatgctact ccgtcaagcc gtcaattgtc tgattcgtta ccaattatga
     4321 caacttgacg gctacatcat tcactttttc ttcacaaccg gcacggaact cgctcgggct
     4381 ggccccggtg cattttttaa atacccgcga gaaatagagt tgatcgtcaa aaccaacatt
     4441 gcgaccgacg gtggcgatag gcatccgggt ggtgctcaaa agcagcttcg cctggctgat
     4501 acgttggtcc tcgcgccagc ttaagacgct aatccctaac tgctggcgga aaagatgtga
     4561 cagacgcgac ggcgacaagc aaacatgctg tgcgacgctg gcgatatcaa aattgctgtc
     4621 tgccaggtga tcgctgatgt actgacaagc ctcgcgtacc cgattatcca tcggtggatg
     4681 gagcgactcg ttaatcgctt ccatgcgccg cagtaacaat tgctcaagca gatttatcgc
     4741 cagcagctcc gaatagcgcc cttccccttg cccggcgtta atgatttgcc caaacaggtc
     4801 gctgaaatgc ggctggtgcg cttcatccgg gcgaaagaac cccgtattgg caaatattga
     4861 cggccagtta agccattcat gccagtaggc gcgcggacga aagtaaaccc actggtgata
     4921 ccattcgcga gcctccggat gacgaccgta gtgatgaatc tctcctggcg ggaacagcaa
     4981 aatatcaccc ggtcggcaaa caaattctcg tccctgattt ttcaccaccc cctgaccgcg
     5041 aatggtgaga ttgagaatat aacctttcat tcccagcggt cggtcgataa aaaaatcgag
     5101 ataaccgttg gcctcaatcg gcgttaaacc cgccaccaga tgggcattaa acgagtatcc
     5161 cggcagcagg ggatcatttt gcgcttcagc catacttttc atactcccgc cattcagaga
     5221 agaaaccaat tgtccatatt gcatcagaca ttgccgtcac tgcgtctttt actggctctt
     5281 ctcgctaacc aaaccggtaa ccccgcttat taaaagcatt ctgtaacaaa gcgggaccaa
     5341 agccatgaca aaaacgcgta acaaaagtgt ctataatcac ggcagaaaag tccacattga
     5401 ttatttgcac ggcgtcacac tttgctatgc catagcattt ttatccataa gattagcgga
     5461 tcctacctga cgctttttat cgcaactctc tactgtttct ccatacgtcg acagctagcg
     5521 tattttggat gataaggagg cgcaaaaaat ggcgagcagt gaggacgtca tcaaggagtt
     5581 catgcgcttc aaggtgcgca tggagggctc cgtgaacggc cacgagttcg agatcgaggg
     5641 cgagggcgag ggccgcccct acgagggcac ccagaccgcc aagctgaagg tgaccaaggg
     5701 cggccccctg cccttcgcct gggacatcct gtccccccag ttccagtacg gctccaaggt
     5761 gtacgtgaag caccccgccg acatccccga ctacaagaag ctgtccttcc ccgagggctt
     5821 caagtgggag cgcgtgatga acttcgagga cggcggcgtg gtgaccgtga cccaggactc
     5881 ctccctgcag gacggctcct tcatctacaa ggtgaagttc atcggcgtga acttcccctc
     5941 cgacggcccc gtaatgcaga agaagactat gggctgggag gcctccaccg agcgcctgta
     6001 cccccgcgac ggcgtgctga agggcgagat ccacaaggcc ctgaagctga aggacggcgg
     6061 ccactacctg gtggagttca agtccatcta catggccaag aagcccgtgc agctgcccgg
     6121 ctactactac gtggactcca agctggacat cacctcccac aacgaggact acaccatcgt
     6181 ggagcagtac gagcgcgccg agggccgcca ccacctgttc ctgta
//
