LOCUS       sequence-64692-e        5110 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 5110)
  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 5110)
  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-L3S2P00
FEATURES             Location/Qualifiers
     source          1..5110
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             90..101
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             108..785
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     terminator      830..901
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      917..944
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     primer_bind     complement(1038..1055)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1209..1797)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1289..1308)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1968..2759,2760..2828))
                     /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     2591..2610
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2829..2933)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3001..3019
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(3023..3066)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     CDS             complement(3155..4033)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        4060..4344
                     /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     4267..4286
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     RBS             4373..4384
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             4391..5104
                     /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(4421..4449)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     5044..5064
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
ORIGIN
        1 gaattcctcg gtaccaaatt ccagaaaaga ggggagcggg aaaccgctcc ccttttttcg
       61 ttttggtcca ctagtagcgg ccgctgcaga aagaggagaa atactagatg gcttcctccg
      121 aagacgttat caaagagttc atgcgtttca aagttcgtat ggaaggttcc gttaacggtc
      181 acgagttcga aatcgaaggt gaaggtgaag gtcgtccgta cgaaggtacc cagaccgcta
      241 aactgaaagt taccaaaggt ggtccgctgc cgttcgcttg ggacatcctg tccccgcagt
      301 tccagtacgg ttccaaagct tacgttaaac acccggctga catcccggac tacctgaaac
      361 tgtccttccc ggaaggtttc aaatgggaac gtgttatgaa cttcgaagac ggtggtgttg
      421 ttaccgttac ccaggactcc tccctgcaag acggtgagtt catctacaaa gttaaactgc
      481 gtggtaccaa cttcccgtcc gacggtccgg ttatgcagaa aaaaaccatg ggttgggaag
      541 cttccaccga acgtatgtac ccggaagacg gtgctctgaa aggtgaaatc aaaatgcgtc
      601 tgaaactgaa agacggtggt cactacgacg ctgaagttaa aaccacctac atggctaaaa
      661 aaccggttca gctgccgggt gcttacaaaa ccgacatcaa actggacatc acctcccaca
      721 acgaagacta caccatcgtt gaacagtacg aacgtgctga aggtcgtcac tccaccggtg
      781 cttaataacg ctgatagtgc tagtgtagat cgctactaga gccaggcatc aaataaaacg
      841 aaaggctcag tcgaaagact gggcctttcg ttttatctgt tgtttgtcgg tgaacgctct
      901 ctactagagt cacactggct caccttcggg tgggcctttc tgcgtttata tactagagtc
      961 aattgaggta gaggtacaca cgcgaactcc gatagccaat tcagagtaat aaactgtgat
     1021 aatcaatgca gtgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     1081 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     1141 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     1201 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     1261 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     1321 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     1381 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     1441 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     1501 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     1561 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     1621 tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag
     1681 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     1741 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     1801 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     1861 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     1921 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     1981 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     2041 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     2101 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     2161 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     2221 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     2281 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     2341 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     2401 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     2461 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     2521 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     2581 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     2641 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     2701 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     2761 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     2821 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     2881 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     2941 ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa
     3001 aataggcgta tcacgaggca gaatttcaga taaaaaaaat ccttagcttt cgctaaggat
     3061 gatttctgga attgaattga ggtagaggta cacacgcgaa ctccgatagc caattcagag
     3121 taataaactg tgataatcaa tgcatgtact agagttatga caacttgacg gctacatcat
     3181 tcactttttc ttcacaaccg gcacggaact cgctcgggct ggccccggtg cattttttaa
     3241 atacccgcga gaaatagagt tgatcgtcaa aaccaacatt gcgaccgacg gtggcgatag
     3301 gcatccgggt ggtgctcaaa agcagcttcg cctggctgat acgttggtcc tcgcgccagc
     3361 ttaagacgct aatccctaac tgctggcgga aaagatgtga cagacgcgac ggcgacaagc
     3421 aaacatgctg tgcgacgctg gcgatatcaa aattgctgtc tgccaggtga tcgctgatgt
     3481 actgacaagc ctcgcgtacc cgattatcca tcggtggatg gagcgactcg ttaatcgctt
     3541 ccatgcgccg cagtaacaat tgctcaagca gatttatcgc cagcagctcc gaatagcgcc
     3601 cttccccttg cccggcgtta atgatttgcc caaacaggtc gctgaaatgc ggctggtgcg
     3661 cttcatccgg gcgaaagaac cccgtattgg caaatattga cggccagtta agccattcat
     3721 gccagtaggc gcgcggacga aagtaaaccc actggtgata ccattcgcga gcctccggat
     3781 gacgaccgta gtgatgaatc tctcctggcg ggaacagcaa aatatcaccc ggtcggcaaa
     3841 caaattctcg tccctgattt ttcaccaccc cctgaccgcg aatggtgaga ttgagaatat
     3901 aacctttcat tcccagcggt cggtcgataa aaaaatcgag ataaccgttg gcctcaatcg
     3961 gcgttaaacc cgccaccaga tgggcattaa acgagtatcc cggcagcagg ggatcatttt
     4021 gcgcttcagc catacttttc atactcccgc cattcagaga agaaaccaat tgtccatatt
     4081 gcatcagaca ttgccgtcac tgcgtctttt actggctctt ctcgctaacc aaaccggtaa
     4141 ccccgcttat taaaagcatt ctgtaacaaa gcgggaccaa agccatgaca aaaacgcgta
     4201 acaaaagtgt ctataatcac ggcagaaaag tccacattga ttatttgcac ggcgtcacac
     4261 tttgctatgc catagcattt ttatccataa gattagcgga tcctacctga cgctttttat
     4321 cgcaactctc tactgtttct ccatacccgt ttttttgggc tagctactag agaaagagga
     4381 gaaatactag atgcgtaaag gagaagaact tttcactgga gttgtcccaa ttcttgttga
     4441 attagatggt gatgttaatg ggcacaaatt ttctgtcagt ggagagggtg aaggtgatgc
     4501 aacatacgga aaacttaccc ttaaatttat ttgcactact ggaaaactac ctgttccatg
     4561 gccaacactt gtcactactt tcggttatgg tgttcaatgc tttgcgagat acccagatca
     4621 tatgaaacag catgactttt tcaagagtgc catgcccgaa ggttatgtac aggaaagaac
     4681 tatatttttc aaagatgacg ggaactacaa gacacgtgct gaagtcaagt ttgaaggtga
     4741 tacccttgtt aatagaatcg agttaaaagg tattgatttt aaagaagatg gaaacattct
     4801 tggacacaaa ttggaataca actataactc acacaatgta tacatcatgg cagacaaaca
     4861 aaagaatgga atcaaagtta acttcaaaat tagacacaac attgaagatg gaagcgttca
     4921 actagcagac cattatcaac aaaatactcc aattggcgat ggccctgtcc ttttaccaga
     4981 caaccattac ctgtccacac aatctgccct ttcgaaagat cccaacgaaa agagagacca
     5041 catggtcctt cttgagtttg taacagctgc tgggattaca catggcatgg atgaactata
     5101 caaataataa
//
