LOCUS       sequence-64700-e        5096 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 5096)
  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 5096)
  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-ECK120010783
FEATURES             Location/Qualifiers
     source          1..5096
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             76..87
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             94..771
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     terminator      816..887
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      903..930
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     primer_bind     complement(1024..1041)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1195..1783)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1275..1294)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1954..2745,2746..2814))
                     /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     2577..2596
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2815..2919)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2987..3005
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(3009..3052)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     CDS             complement(3141..4019)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        4046..4330
                     /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     4253..4272
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     RBS             4359..4370
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             4377..5090
                     /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(4407..4435)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     5030..5050
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
ORIGIN
        1 gaattcacga gccaataaaa ataccggcgt tatgccggta tttttttacg aaagaactag
       61 tagcggccgc tgcagaaaga ggagaaatac tagatggctt cctccgaaga cgttatcaaa
      121 gagttcatgc gtttcaaagt tcgtatggaa ggttccgtta acggtcacga gttcgaaatc
      181 gaaggtgaag gtgaaggtcg tccgtacgaa ggtacccaga ccgctaaact gaaagttacc
      241 aaaggtggtc cgctgccgtt cgcttgggac atcctgtccc cgcagttcca gtacggttcc
      301 aaagcttacg ttaaacaccc ggctgacatc ccggactacc tgaaactgtc cttcccggaa
      361 ggtttcaaat gggaacgtgt tatgaacttc gaagacggtg gtgttgttac cgttacccag
      421 gactcctccc tgcaagacgg tgagttcatc tacaaagtta aactgcgtgg taccaacttc
      481 ccgtccgacg gtccggttat gcagaaaaaa accatgggtt gggaagcttc caccgaacgt
      541 atgtacccgg aagacggtgc tctgaaaggt gaaatcaaaa tgcgtctgaa actgaaagac
      601 ggtggtcact acgacgctga agttaaaacc acctacatgg ctaaaaaacc ggttcagctg
      661 ccgggtgctt acaaaaccga catcaaactg gacatcacct cccacaacga agactacacc
      721 atcgttgaac agtacgaacg tgctgaaggt cgtcactcca ccggtgctta ataacgctga
      781 tagtgctagt gtagatcgct actagagcca ggcatcaaat aaaacgaaag gctcagtcga
      841 aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa cgctctctac tagagtcaca
      901 ctggctcacc ttcgggtggg cctttctgcg tttatatact agagtcaatt gaggtagagg
      961 tacacacgcg aactccgata gccaattcag agtaataaac tgtgataatc aatgcagtgc
     1021 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     1081 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     1141 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     1201 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     1261 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     1321 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     1381 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     1441 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     1501 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     1561 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     1621 cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     1681 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     1741 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     1801 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     1861 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     1921 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     1981 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     2041 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     2101 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     2161 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     2221 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     2281 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     2341 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     2401 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     2461 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     2521 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     2581 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     2641 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     2701 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     2761 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     2821 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     2881 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     2941 acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac
     3001 gaggcagaat ttcagataaa aaaaatcctt agctttcgct aaggatgatt tctggaattg
     3061 aattgaggta gaggtacaca cgcgaactcc gatagccaat tcagagtaat aaactgtgat
     3121 aatcaatgca tgtactagag ttatgacaac ttgacggcta catcattcac tttttcttca
     3181 caaccggcac ggaactcgct cgggctggcc ccggtgcatt ttttaaatac ccgcgagaaa
     3241 tagagttgat cgtcaaaacc aacattgcga ccgacggtgg cgataggcat ccgggtggtg
     3301 ctcaaaagca gcttcgcctg gctgatacgt tggtcctcgc gccagcttaa gacgctaatc
     3361 cctaactgct ggcggaaaag atgtgacaga cgcgacggcg acaagcaaac atgctgtgcg
     3421 acgctggcga tatcaaaatt gctgtctgcc aggtgatcgc tgatgtactg acaagcctcg
     3481 cgtacccgat tatccatcgg tggatggagc gactcgttaa tcgcttccat gcgccgcagt
     3541 aacaattgct caagcagatt tatcgccagc agctccgaat agcgcccttc cccttgcccg
     3601 gcgttaatga tttgcccaaa caggtcgctg aaatgcggct ggtgcgcttc atccgggcga
     3661 aagaaccccg tattggcaaa tattgacggc cagttaagcc attcatgcca gtaggcgcgc
     3721 ggacgaaagt aaacccactg gtgataccat tcgcgagcct ccggatgacg accgtagtga
     3781 tgaatctctc ctggcgggaa cagcaaaata tcacccggtc ggcaaacaaa ttctcgtccc
     3841 tgatttttca ccaccccctg accgcgaatg gtgagattga gaatataacc tttcattccc
     3901 agcggtcggt cgataaaaaa atcgagataa ccgttggcct caatcggcgt taaacccgcc
     3961 accagatggg cattaaacga gtatcccggc agcaggggat cattttgcgc ttcagccata
     4021 cttttcatac tcccgccatt cagagaagaa accaattgtc catattgcat cagacattgc
     4081 cgtcactgcg tcttttactg gctcttctcg ctaaccaaac cggtaacccc gcttattaaa
     4141 agcattctgt aacaaagcgg gaccaaagcc atgacaaaaa cgcgtaacaa aagtgtctat
     4201 aatcacggca gaaaagtcca cattgattat ttgcacggcg tcacactttg ctatgccata
     4261 gcatttttat ccataagatt agcggatcct acctgacgct ttttatcgca actctctact
     4321 gtttctccat acccgttttt ttgggctagc tactagagaa agaggagaaa tactagatgc
     4381 gtaaaggaga agaacttttc actggagttg tcccaattct tgttgaatta gatggtgatg
     4441 ttaatgggca caaattttct gtcagtggag agggtgaagg tgatgcaaca tacggaaaac
     4501 ttacccttaa atttatttgc actactggaa aactacctgt tccatggcca acacttgtca
     4561 ctactttcgg ttatggtgtt caatgctttg cgagataccc agatcatatg aaacagcatg
     4621 actttttcaa gagtgccatg cccgaaggtt atgtacagga aagaactata tttttcaaag
     4681 atgacgggaa ctacaagaca cgtgctgaag tcaagtttga aggtgatacc cttgttaata
     4741 gaatcgagtt aaaaggtatt gattttaaag aagatggaaa cattcttgga cacaaattgg
     4801 aatacaacta taactcacac aatgtataca tcatggcaga caaacaaaag aatggaatca
     4861 aagttaactt caaaattaga cacaacattg aagatggaag cgttcaacta gcagaccatt
     4921 atcaacaaaa tactccaatt ggcgatggcc ctgtcctttt accagacaac cattacctgt
     4981 ccacacaatc tgccctttcg aaagatccca acgaaaagag agaccacatg gtccttcttg
     5041 agtttgtaac agctgctggg attacacatg gcatggatga actatacaaa taataa
//
