LOCUS       sequence-64697-e        5104 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 5104)
  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 5104)
  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-L3S2P36
FEATURES             Location/Qualifiers
     source          1..5104
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             84..95
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             102..779
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     terminator      824..895
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      911..938
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     primer_bind     complement(1032..1049)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1203..1791)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1283..1302)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1962..2753,2754..2822))
                     /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     2585..2604
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2823..2927)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2995..3013
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(3017..3060)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     CDS             complement(3149..4027)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        4054..4338
                     /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     4261..4280
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     RBS             4367..4378
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             4385..5098
                     /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(4415..4443)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     5038..5058
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
ORIGIN
        1 gaattcctcg gtaccaaatt ccagaaaaga gacgctgaaa agcgtctttt ttattgatgg
       61 tccactagta gcggccgctg cagaaagagg agaaatacta gatggcttcc tccgaagacg
      121 ttatcaaaga gttcatgcgt ttcaaagttc gtatggaagg ttccgttaac ggtcacgagt
      181 tcgaaatcga aggtgaaggt gaaggtcgtc cgtacgaagg tacccagacc gctaaactga
      241 aagttaccaa aggtggtccg ctgccgttcg cttgggacat cctgtccccg cagttccagt
      301 acggttccaa agcttacgtt aaacacccgg ctgacatccc ggactacctg aaactgtcct
      361 tcccggaagg tttcaaatgg gaacgtgtta tgaacttcga agacggtggt gttgttaccg
      421 ttacccagga ctcctccctg caagacggtg agttcatcta caaagttaaa ctgcgtggta
      481 ccaacttccc gtccgacggt ccggttatgc agaaaaaaac catgggttgg gaagcttcca
      541 ccgaacgtat gtacccggaa gacggtgctc tgaaaggtga aatcaaaatg cgtctgaaac
      601 tgaaagacgg tggtcactac gacgctgaag ttaaaaccac ctacatggct aaaaaaccgg
      661 ttcagctgcc gggtgcttac aaaaccgaca tcaaactgga catcacctcc cacaacgaag
      721 actacaccat cgttgaacag tacgaacgtg ctgaaggtcg tcactccacc ggtgcttaat
      781 aacgctgata gtgctagtgt agatcgctac tagagccagg catcaaataa aacgaaaggc
      841 tcagtcgaaa gactgggcct ttcgttttat ctgttgtttg tcggtgaacg ctctctacta
      901 gagtcacact ggctcacctt cgggtgggcc tttctgcgtt tatatactag agtcaattga
      961 ggtagaggta cacacgcgaa ctccgatagc caattcagag taataaactg tgataatcaa
     1021 tgcagtgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     1081 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     1141 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     1201 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     1261 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     1321 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     1381 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     1441 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     1501 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     1561 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     1621 cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt
     1681 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     1741 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     1801 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     1861 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     1921 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     1981 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     2041 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     2101 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     2161 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     2221 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     2281 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     2341 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     2401 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     2461 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     2521 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     2581 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     2641 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     2701 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     2761 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     2821 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     2881 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     2941 aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg
     3001 cgtatcacga ggcagaattt cagataaaaa aaatccttag ctttcgctaa ggatgatttc
     3061 tggaattgaa ttgaggtaga ggtacacacg cgaactccga tagccaattc agagtaataa
     3121 actgtgataa tcaatgcatg tactagagtt atgacaactt gacggctaca tcattcactt
     3181 tttcttcaca accggcacgg aactcgctcg ggctggcccc ggtgcatttt ttaaataccc
     3241 gcgagaaata gagttgatcg tcaaaaccaa cattgcgacc gacggtggcg ataggcatcc
     3301 gggtggtgct caaaagcagc ttcgcctggc tgatacgttg gtcctcgcgc cagcttaaga
     3361 cgctaatccc taactgctgg cggaaaagat gtgacagacg cgacggcgac aagcaaacat
     3421 gctgtgcgac gctggcgata tcaaaattgc tgtctgccag gtgatcgctg atgtactgac
     3481 aagcctcgcg tacccgatta tccatcggtg gatggagcga ctcgttaatc gcttccatgc
     3541 gccgcagtaa caattgctca agcagattta tcgccagcag ctccgaatag cgcccttccc
     3601 cttgcccggc gttaatgatt tgcccaaaca ggtcgctgaa atgcggctgg tgcgcttcat
     3661 ccgggcgaaa gaaccccgta ttggcaaata ttgacggcca gttaagccat tcatgccagt
     3721 aggcgcgcgg acgaaagtaa acccactggt gataccattc gcgagcctcc ggatgacgac
     3781 cgtagtgatg aatctctcct ggcgggaaca gcaaaatatc acccggtcgg caaacaaatt
     3841 ctcgtccctg atttttcacc accccctgac cgcgaatggt gagattgaga atataacctt
     3901 tcattcccag cggtcggtcg ataaaaaaat cgagataacc gttggcctca atcggcgtta
     3961 aacccgccac cagatgggca ttaaacgagt atcccggcag caggggatca ttttgcgctt
     4021 cagccatact tttcatactc ccgccattca gagaagaaac caattgtcca tattgcatca
     4081 gacattgccg tcactgcgtc ttttactggc tcttctcgct aaccaaaccg gtaaccccgc
     4141 ttattaaaag cattctgtaa caaagcggga ccaaagccat gacaaaaacg cgtaacaaaa
     4201 gtgtctataa tcacggcaga aaagtccaca ttgattattt gcacggcgtc acactttgct
     4261 atgccatagc atttttatcc ataagattag cggatcctac ctgacgcttt ttatcgcaac
     4321 tctctactgt ttctccatac ccgttttttt gggctagcta ctagagaaag aggagaaata
     4381 ctagatgcgt aaaggagaag aacttttcac tggagttgtc ccaattcttg ttgaattaga
     4441 tggtgatgtt aatgggcaca aattttctgt cagtggagag ggtgaaggtg atgcaacata
     4501 cggaaaactt acccttaaat ttatttgcac tactggaaaa ctacctgttc catggccaac
     4561 acttgtcact actttcggtt atggtgttca atgctttgcg agatacccag atcatatgaa
     4621 acagcatgac tttttcaaga gtgccatgcc cgaaggttat gtacaggaaa gaactatatt
     4681 tttcaaagat gacgggaact acaagacacg tgctgaagtc aagtttgaag gtgataccct
     4741 tgttaataga atcgagttaa aaggtattga ttttaaagaa gatggaaaca ttcttggaca
     4801 caaattggaa tacaactata actcacacaa tgtatacatc atggcagaca aacaaaagaa
     4861 tggaatcaaa gttaacttca aaattagaca caacattgaa gatggaagcg ttcaactagc
     4921 agaccattat caacaaaata ctccaattgg cgatggccct gtccttttac cagacaacca
     4981 ttacctgtcc acacaatctg ccctttcgaa agatcccaac gaaaagagag accacatggt
     5041 ccttcttgag tttgtaacag ctgctgggat tacacatggc atggatgaac tatacaaata
     5101 ataa
//
