LOCUS       sequence-64696-e        5094 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 5094)
  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 5094)
  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-ECK120015170
FEATURES             Location/Qualifiers
     source          1..5094
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             74..85
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             92..769
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     terminator      814..885
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      901..928
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     primer_bind     complement(1022..1039)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1193..1781)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1273..1292)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1952..2743,2744..2812))
                     /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     2575..2594
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2813..2917)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2985..3003
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(3007..3050)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     CDS             complement(3139..4017)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        4044..4328
                     /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     4251..4270
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     RBS             4357..4368
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             4375..5088
                     /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(4405..4433)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     5028..5048
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
ORIGIN
        1 gaattcacaa ttttcgaaaa aacccgcttc ggcgggtttt tttatagcta aaaactagta
       61 gcggccgctg cagaaagagg agaaatacta gatggcttcc tccgaagacg ttatcaaaga
      121 gttcatgcgt ttcaaagttc gtatggaagg ttccgttaac ggtcacgagt tcgaaatcga
      181 aggtgaaggt gaaggtcgtc cgtacgaagg tacccagacc gctaaactga aagttaccaa
      241 aggtggtccg ctgccgttcg cttgggacat cctgtccccg cagttccagt acggttccaa
      301 agcttacgtt aaacacccgg ctgacatccc ggactacctg aaactgtcct tcccggaagg
      361 tttcaaatgg gaacgtgtta tgaacttcga agacggtggt gttgttaccg ttacccagga
      421 ctcctccctg caagacggtg agttcatcta caaagttaaa ctgcgtggta ccaacttccc
      481 gtccgacggt ccggttatgc agaaaaaaac catgggttgg gaagcttcca ccgaacgtat
      541 gtacccggaa gacggtgctc tgaaaggtga aatcaaaatg cgtctgaaac tgaaagacgg
      601 tggtcactac gacgctgaag ttaaaaccac ctacatggct aaaaaaccgg ttcagctgcc
      661 gggtgcttac aaaaccgaca tcaaactgga catcacctcc cacaacgaag actacaccat
      721 cgttgaacag tacgaacgtg ctgaaggtcg tcactccacc ggtgcttaat aacgctgata
      781 gtgctagtgt agatcgctac tagagccagg catcaaataa aacgaaaggc tcagtcgaaa
      841 gactgggcct ttcgttttat ctgttgtttg tcggtgaacg ctctctacta gagtcacact
      901 ggctcacctt cgggtgggcc tttctgcgtt tatatactag agtcaattga ggtagaggta
      961 cacacgcgaa ctccgatagc caattcagag taataaactg tgataatcaa tgcagtgctt
     1021 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     1081 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     1141 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     1201 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     1261 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     1321 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     1381 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     1441 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     1501 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     1561 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     1621 gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     1681 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     1741 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     1801 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     1861 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     1921 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     1981 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     2041 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     2101 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     2161 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     2221 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     2281 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     2341 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     2401 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     2461 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     2521 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     2581 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     2641 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     2701 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     2761 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     2821 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     2881 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     2941 ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga
     3001 ggcagaattt cagataaaaa aaatccttag ctttcgctaa ggatgatttc tggaattgaa
     3061 ttgaggtaga ggtacacacg cgaactccga tagccaattc agagtaataa actgtgataa
     3121 tcaatgcatg tactagagtt atgacaactt gacggctaca tcattcactt tttcttcaca
     3181 accggcacgg aactcgctcg ggctggcccc ggtgcatttt ttaaataccc gcgagaaata
     3241 gagttgatcg tcaaaaccaa cattgcgacc gacggtggcg ataggcatcc gggtggtgct
     3301 caaaagcagc ttcgcctggc tgatacgttg gtcctcgcgc cagcttaaga cgctaatccc
     3361 taactgctgg cggaaaagat gtgacagacg cgacggcgac aagcaaacat gctgtgcgac
     3421 gctggcgata tcaaaattgc tgtctgccag gtgatcgctg atgtactgac aagcctcgcg
     3481 tacccgatta tccatcggtg gatggagcga ctcgttaatc gcttccatgc gccgcagtaa
     3541 caattgctca agcagattta tcgccagcag ctccgaatag cgcccttccc cttgcccggc
     3601 gttaatgatt tgcccaaaca ggtcgctgaa atgcggctgg tgcgcttcat ccgggcgaaa
     3661 gaaccccgta ttggcaaata ttgacggcca gttaagccat tcatgccagt aggcgcgcgg
     3721 acgaaagtaa acccactggt gataccattc gcgagcctcc ggatgacgac cgtagtgatg
     3781 aatctctcct ggcgggaaca gcaaaatatc acccggtcgg caaacaaatt ctcgtccctg
     3841 atttttcacc accccctgac cgcgaatggt gagattgaga atataacctt tcattcccag
     3901 cggtcggtcg ataaaaaaat cgagataacc gttggcctca atcggcgtta aacccgccac
     3961 cagatgggca ttaaacgagt atcccggcag caggggatca ttttgcgctt cagccatact
     4021 tttcatactc ccgccattca gagaagaaac caattgtcca tattgcatca gacattgccg
     4081 tcactgcgtc ttttactggc tcttctcgct aaccaaaccg gtaaccccgc ttattaaaag
     4141 cattctgtaa caaagcggga ccaaagccat gacaaaaacg cgtaacaaaa gtgtctataa
     4201 tcacggcaga aaagtccaca ttgattattt gcacggcgtc acactttgct atgccatagc
     4261 atttttatcc ataagattag cggatcctac ctgacgcttt ttatcgcaac tctctactgt
     4321 ttctccatac ccgttttttt gggctagcta ctagagaaag aggagaaata ctagatgcgt
     4381 aaaggagaag aacttttcac tggagttgtc ccaattcttg ttgaattaga tggtgatgtt
     4441 aatgggcaca aattttctgt cagtggagag ggtgaaggtg atgcaacata cggaaaactt
     4501 acccttaaat ttatttgcac tactggaaaa ctacctgttc catggccaac acttgtcact
     4561 actttcggtt atggtgttca atgctttgcg agatacccag atcatatgaa acagcatgac
     4621 tttttcaaga gtgccatgcc cgaaggttat gtacaggaaa gaactatatt tttcaaagat
     4681 gacgggaact acaagacacg tgctgaagtc aagtttgaag gtgataccct tgttaataga
     4741 atcgagttaa aaggtattga ttttaaagaa gatggaaaca ttcttggaca caaattggaa
     4801 tacaactata actcacacaa tgtatacatc atggcagaca aacaaaagaa tggaatcaaa
     4861 gttaacttca aaattagaca caacattgaa gatggaagcg ttcaactagc agaccattat
     4921 caacaaaata ctccaattgg cgatggccct gtccttttac cagacaacca ttacctgtcc
     4981 acacaatctg ccctttcgaa agatcccaac gaaaagagag accacatggt ccttcttgag
     5041 tttgtaacag ctgctgggat tacacatggc atggatgaac tatacaaata ataa
//
