LOCUS       sequence-64698-e        5125 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 5125)
  AUTHORS   Chen YJ, Liu P, Nielsen AA, Brophy JA, Clancy K, Peterson T, Voigt 
            CA
  TITLE     Characterization of 582 natural and synthetic terminators and 
            quantification of their design constraints.
  JOURNAL   Nat Methods. 2013 Jun 2. doi: 10.1038/nmeth.2515.
  PUBMED    23727987
REFERENCE   2  (bases 1 to 5125)
  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: "Nat 
            Methods. 2013 Jun 2. doi: 10.1038/nmeth.2515."
COMMENT     Sequence Label: pGR-ECK125108723
FEATURES             Location/Qualifiers
     source          1..5125
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             105..116
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             123..800
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     terminator      845..916
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      932..959
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     primer_bind     complement(1053..1070)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1224..1812)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1304..1323)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1983..2774,2775..2843))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     2606..2625
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2844..2948)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3016..3034
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(3038..3081)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     CDS             complement(3170..4048)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        4075..4359
                     /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     4282..4301
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     RBS             4388..4399
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             4406..5119
                     /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(4436..4464)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     5059..5079
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
ORIGIN
        1 gaattctagc gtaaaagcaa aacacaaatc tatccatgca agcattcacc gccggtttac
       61 tggcggtttt ttttcgccgt cataactagt agcggccgct gcagaaagag gagaaatact
      121 agatggcttc ctccgaagac gttatcaaag agttcatgcg tttcaaagtt cgtatggaag
      181 gttccgttaa cggtcacgag ttcgaaatcg aaggtgaagg tgaaggtcgt ccgtacgaag
      241 gtacccagac cgctaaactg aaagttacca aaggtggtcc gctgccgttc gcttgggaca
      301 tcctgtcccc gcagttccag tacggttcca aagcttacgt taaacacccg gctgacatcc
      361 cggactacct gaaactgtcc ttcccggaag gtttcaaatg ggaacgtgtt atgaacttcg
      421 aagacggtgg tgttgttacc gttacccagg actcctccct gcaagacggt gagttcatct
      481 acaaagttaa actgcgtggt accaacttcc cgtccgacgg tccggttatg cagaaaaaaa
      541 ccatgggttg ggaagcttcc accgaacgta tgtacccgga agacggtgct ctgaaaggtg
      601 aaatcaaaat gcgtctgaaa ctgaaagacg gtggtcacta cgacgctgaa gttaaaacca
      661 cctacatggc taaaaaaccg gttcagctgc cgggtgctta caaaaccgac atcaaactgg
      721 acatcacctc ccacaacgaa gactacacca tcgttgaaca gtacgaacgt gctgaaggtc
      781 gtcactccac cggtgcttaa taacgctgat agtgctagtg tagatcgcta ctagagccag
      841 gcatcaaata aaacgaaagg ctcagtcgaa agactgggcc tttcgtttta tctgttgttt
      901 gtcggtgaac gctctctact agagtcacac tggctcacct tcgggtgggc ctttctgcgt
      961 ttatatacta gagtcaattg aggtagaggt acacacgcga actccgatag ccaattcaga
     1021 gtaataaact gtgataatca atgcagtgct tcctcgctca ctgactcgct gcgctcggtc
     1081 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     1141 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     1201 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     1261 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     1321 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     1381 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     1441 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     1501 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     1561 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     1621 acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc
     1681 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     1741 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     1801 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     1861 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     1921 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     1981 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     2041 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     2101 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     2161 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     2221 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     2281 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     2341 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     2401 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     2461 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     2521 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     2581 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     2641 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     2701 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     2761 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     2821 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     2881 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     2941 gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg
     3001 acattaacct ataaaaatag gcgtatcacg aggcagaatt tcagataaaa aaaatcctta
     3061 gctttcgcta aggatgattt ctggaattga attgaggtag aggtacacac gcgaactccg
     3121 atagccaatt cagagtaata aactgtgata atcaatgcat gtactagagt tatgacaact
     3181 tgacggctac atcattcact ttttcttcac aaccggcacg gaactcgctc gggctggccc
     3241 cggtgcattt tttaaatacc cgcgagaaat agagttgatc gtcaaaacca acattgcgac
     3301 cgacggtggc gataggcatc cgggtggtgc tcaaaagcag cttcgcctgg ctgatacgtt
     3361 ggtcctcgcg ccagcttaag acgctaatcc ctaactgctg gcggaaaaga tgtgacagac
     3421 gcgacggcga caagcaaaca tgctgtgcga cgctggcgat atcaaaattg ctgtctgcca
     3481 ggtgatcgct gatgtactga caagcctcgc gtacccgatt atccatcggt ggatggagcg
     3541 actcgttaat cgcttccatg cgccgcagta acaattgctc aagcagattt atcgccagca
     3601 gctccgaata gcgcccttcc ccttgcccgg cgttaatgat ttgcccaaac aggtcgctga
     3661 aatgcggctg gtgcgcttca tccgggcgaa agaaccccgt attggcaaat attgacggcc
     3721 agttaagcca ttcatgccag taggcgcgcg gacgaaagta aacccactgg tgataccatt
     3781 cgcgagcctc cggatgacga ccgtagtgat gaatctctcc tggcgggaac agcaaaatat
     3841 cacccggtcg gcaaacaaat tctcgtccct gatttttcac caccccctga ccgcgaatgg
     3901 tgagattgag aatataacct ttcattccca gcggtcggtc gataaaaaaa tcgagataac
     3961 cgttggcctc aatcggcgtt aaacccgcca ccagatgggc attaaacgag tatcccggca
     4021 gcaggggatc attttgcgct tcagccatac ttttcatact cccgccattc agagaagaaa
     4081 ccaattgtcc atattgcatc agacattgcc gtcactgcgt cttttactgg ctcttctcgc
     4141 taaccaaacc ggtaaccccg cttattaaaa gcattctgta acaaagcggg accaaagcca
     4201 tgacaaaaac gcgtaacaaa agtgtctata atcacggcag aaaagtccac attgattatt
     4261 tgcacggcgt cacactttgc tatgccatag catttttatc cataagatta gcggatccta
     4321 cctgacgctt tttatcgcaa ctctctactg tttctccata cccgtttttt tgggctagct
     4381 actagagaaa gaggagaaat actagatgcg taaaggagaa gaacttttca ctggagttgt
     4441 cccaattctt gttgaattag atggtgatgt taatgggcac aaattttctg tcagtggaga
     4501 gggtgaaggt gatgcaacat acggaaaact tacccttaaa tttatttgca ctactggaaa
     4561 actacctgtt ccatggccaa cacttgtcac tactttcggt tatggtgttc aatgctttgc
     4621 gagataccca gatcatatga aacagcatga ctttttcaag agtgccatgc ccgaaggtta
     4681 tgtacaggaa agaactatat ttttcaaaga tgacgggaac tacaagacac gtgctgaagt
     4741 caagtttgaa ggtgataccc ttgttaatag aatcgagtta aaaggtattg attttaaaga
     4801 agatggaaac attcttggac acaaattgga atacaactat aactcacaca atgtatacat
     4861 catggcagac aaacaaaaga atggaatcaa agttaacttc aaaattagac acaacattga
     4921 agatggaagc gttcaactag cagaccatta tcaacaaaat actccaattg gcgatggccc
     4981 tgtcctttta ccagacaacc attacctgtc cacacaatct gccctttcga aagatcccaa
     5041 cgaaaagaga gaccacatgg tccttcttga gtttgtaaca gctgctggga ttacacatgg
     5101 catggatgaa ctatacaaat aataa
//
