LOCUS       sequence-64702-e        5092 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 5092)
  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 5092)
  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-ECK120033127
FEATURES             Location/Qualifiers
     source          1..5092
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             72..83
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             90..767
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     terminator      812..883
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      899..926
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     primer_bind     complement(1020..1037)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1191..1779)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1271..1290)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1950..2741,2742..2810))
                     /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     2573..2592
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2811..2915)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2983..3001
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(3005..3048)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     CDS             complement(3137..4015)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        4042..4326
                     /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     4249..4268
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     RBS             4355..4366
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             4373..5086
                     /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(4403..4431)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     5026..5046
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
ORIGIN
        1 gaattctact tcttactcgc ccatctgcaa cggatgggcg aatttatacc cactagtagc
       61 ggccgctgca gaaagaggag aaatactaga tggcttcctc cgaagacgtt atcaaagagt
      121 tcatgcgttt caaagttcgt atggaaggtt ccgttaacgg tcacgagttc gaaatcgaag
      181 gtgaaggtga aggtcgtccg tacgaaggta cccagaccgc taaactgaaa gttaccaaag
      241 gtggtccgct gccgttcgct tgggacatcc tgtccccgca gttccagtac ggttccaaag
      301 cttacgttaa acacccggct gacatcccgg actacctgaa actgtccttc ccggaaggtt
      361 tcaaatggga acgtgttatg aacttcgaag acggtggtgt tgttaccgtt acccaggact
      421 cctccctgca agacggtgag ttcatctaca aagttaaact gcgtggtacc aacttcccgt
      481 ccgacggtcc ggttatgcag aaaaaaacca tgggttggga agcttccacc gaacgtatgt
      541 acccggaaga cggtgctctg aaaggtgaaa tcaaaatgcg tctgaaactg aaagacggtg
      601 gtcactacga cgctgaagtt aaaaccacct acatggctaa aaaaccggtt cagctgccgg
      661 gtgcttacaa aaccgacatc aaactggaca tcacctccca caacgaagac tacaccatcg
      721 ttgaacagta cgaacgtgct gaaggtcgtc actccaccgg tgcttaataa cgctgatagt
      781 gctagtgtag atcgctacta gagccaggca tcaaataaaa cgaaaggctc agtcgaaaga
      841 ctgggccttt cgttttatct gttgtttgtc ggtgaacgct ctctactaga gtcacactgg
      901 ctcaccttcg ggtgggcctt tctgcgttta tatactagag tcaattgagg tagaggtaca
      961 cacgcgaact ccgatagcca attcagagta ataaactgtg ataatcaatg cagtgcttcc
     1021 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     1081 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     1141 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     1201 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     1261 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     1321 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     1381 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     1441 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     1501 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     1561 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     1621 tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     1681 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     1741 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     1801 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     1861 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     1921 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     1981 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     2041 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     2101 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     2161 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     2221 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     2281 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     2341 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     2401 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     2461 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     2521 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     2581 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     2641 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     2701 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     2761 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     2821 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     2881 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     2941 gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg
     3001 cagaatttca gataaaaaaa atccttagct ttcgctaagg atgatttctg gaattgaatt
     3061 gaggtagagg tacacacgcg aactccgata gccaattcag agtaataaac tgtgataatc
     3121 aatgcatgta ctagagttat gacaacttga cggctacatc attcactttt tcttcacaac
     3181 cggcacggaa ctcgctcggg ctggccccgg tgcatttttt aaatacccgc gagaaataga
     3241 gttgatcgtc aaaaccaaca ttgcgaccga cggtggcgat aggcatccgg gtggtgctca
     3301 aaagcagctt cgcctggctg atacgttggt cctcgcgcca gcttaagacg ctaatcccta
     3361 actgctggcg gaaaagatgt gacagacgcg acggcgacaa gcaaacatgc tgtgcgacgc
     3421 tggcgatatc aaaattgctg tctgccaggt gatcgctgat gtactgacaa gcctcgcgta
     3481 cccgattatc catcggtgga tggagcgact cgttaatcgc ttccatgcgc cgcagtaaca
     3541 attgctcaag cagatttatc gccagcagct ccgaatagcg cccttcccct tgcccggcgt
     3601 taatgatttg cccaaacagg tcgctgaaat gcggctggtg cgcttcatcc gggcgaaaga
     3661 accccgtatt ggcaaatatt gacggccagt taagccattc atgccagtag gcgcgcggac
     3721 gaaagtaaac ccactggtga taccattcgc gagcctccgg atgacgaccg tagtgatgaa
     3781 tctctcctgg cgggaacagc aaaatatcac ccggtcggca aacaaattct cgtccctgat
     3841 ttttcaccac cccctgaccg cgaatggtga gattgagaat ataacctttc attcccagcg
     3901 gtcggtcgat aaaaaaatcg agataaccgt tggcctcaat cggcgttaaa cccgccacca
     3961 gatgggcatt aaacgagtat cccggcagca ggggatcatt ttgcgcttca gccatacttt
     4021 tcatactccc gccattcaga gaagaaacca attgtccata ttgcatcaga cattgccgtc
     4081 actgcgtctt ttactggctc ttctcgctaa ccaaaccggt aaccccgctt attaaaagca
     4141 ttctgtaaca aagcgggacc aaagccatga caaaaacgcg taacaaaagt gtctataatc
     4201 acggcagaaa agtccacatt gattatttgc acggcgtcac actttgctat gccatagcat
     4261 ttttatccat aagattagcg gatcctacct gacgcttttt atcgcaactc tctactgttt
     4321 ctccataccc gtttttttgg gctagctact agagaaagag gagaaatact agatgcgtaa
     4381 aggagaagaa cttttcactg gagttgtccc aattcttgtt gaattagatg gtgatgttaa
     4441 tgggcacaaa ttttctgtca gtggagaggg tgaaggtgat gcaacatacg gaaaacttac
     4501 ccttaaattt atttgcacta ctggaaaact acctgttcca tggccaacac ttgtcactac
     4561 tttcggttat ggtgttcaat gctttgcgag atacccagat catatgaaac agcatgactt
     4621 tttcaagagt gccatgcccg aaggttatgt acaggaaaga actatatttt tcaaagatga
     4681 cgggaactac aagacacgtg ctgaagtcaa gtttgaaggt gatacccttg ttaatagaat
     4741 cgagttaaaa ggtattgatt ttaaagaaga tggaaacatt cttggacaca aattggaata
     4801 caactataac tcacacaatg tatacatcat ggcagacaaa caaaagaatg gaatcaaagt
     4861 taacttcaaa attagacaca acattgaaga tggaagcgtt caactagcag accattatca
     4921 acaaaatact ccaattggcg atggccctgt ccttttacca gacaaccatt acctgtccac
     4981 acaatctgcc ctttcgaaag atcccaacga aaagagagac cacatggtcc ttcttgagtt
     5041 tgtaacagct gctgggatta cacatggcat ggatgaacta tacaaataat aa
//
