LOCUS       sequence-64686-e        5127 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 5127)
  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 5127)
  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-BBa_B0010
FEATURES             Location/Qualifiers
     source          1..5127
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      15..86
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     RBS             107..118
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             125..802
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     terminator      847..918
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      934..961
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     primer_bind     complement(1055..1072)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1226..1814)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1306..1325)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1985..2776,2777..2845))
                     /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     2608..2627
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2846..2950)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3018..3036
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(3040..3083)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     CDS             complement(3172..4050)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        4077..4361
                     /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     4284..4303
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     RBS             4390..4401
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             4408..5121
                     /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(4438..4466)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     5061..5081
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
ORIGIN
        1 gaattcccag gcatcaaata aaacgaaagg ctcagtcgaa agactgggcc tttcgtttta
       61 tctgttgttt gtcggtgaac gctctcacta gtagcggccg ctgcagaaag aggagaaata
      121 ctagatggct tcctccgaag acgttatcaa agagttcatg cgtttcaaag ttcgtatgga
      181 aggttccgtt aacggtcacg agttcgaaat cgaaggtgaa ggtgaaggtc gtccgtacga
      241 aggtacccag accgctaaac tgaaagttac caaaggtggt ccgctgccgt tcgcttggga
      301 catcctgtcc ccgcagttcc agtacggttc caaagcttac gttaaacacc cggctgacat
      361 cccggactac ctgaaactgt ccttcccgga aggtttcaaa tgggaacgtg ttatgaactt
      421 cgaagacggt ggtgttgtta ccgttaccca ggactcctcc ctgcaagacg gtgagttcat
      481 ctacaaagtt aaactgcgtg gtaccaactt cccgtccgac ggtccggtta tgcagaaaaa
      541 aaccatgggt tgggaagctt ccaccgaacg tatgtacccg gaagacggtg ctctgaaagg
      601 tgaaatcaaa atgcgtctga aactgaaaga cggtggtcac tacgacgctg aagttaaaac
      661 cacctacatg gctaaaaaac cggttcagct gccgggtgct tacaaaaccg acatcaaact
      721 ggacatcacc tcccacaacg aagactacac catcgttgaa cagtacgaac gtgctgaagg
      781 tcgtcactcc accggtgctt aataacgctg atagtgctag tgtagatcgc tactagagcc
      841 aggcatcaaa taaaacgaaa ggctcagtcg aaagactggg cctttcgttt tatctgttgt
      901 ttgtcggtga acgctctcta ctagagtcac actggctcac cttcgggtgg gcctttctgc
      961 gtttatatac tagagtcaat tgaggtagag gtacacacgc gaactccgat agccaattca
     1021 gagtaataaa ctgtgataat caatgcagtg cttcctcgct cactgactcg ctgcgctcgg
     1081 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     1141 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     1201 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     1261 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     1321 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     1381 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     1441 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     1501 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     1561 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     1621 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta
     1681 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     1741 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     1801 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     1861 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     1921 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     1981 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     2041 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     2101 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     2161 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     2221 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     2281 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     2341 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     2401 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     2461 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     2521 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     2581 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     2641 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     2701 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     2761 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     2821 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     2881 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     2941 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca
     3001 tgacattaac ctataaaaat aggcgtatca cgaggcagaa tttcagataa aaaaaatcct
     3061 tagctttcgc taaggatgat ttctggaatt gaattgaggt agaggtacac acgcgaactc
     3121 cgatagccaa ttcagagtaa taaactgtga taatcaatgc atgtactaga gttatgacaa
     3181 cttgacggct acatcattca ctttttcttc acaaccggca cggaactcgc tcgggctggc
     3241 cccggtgcat tttttaaata cccgcgagaa atagagttga tcgtcaaaac caacattgcg
     3301 accgacggtg gcgataggca tccgggtggt gctcaaaagc agcttcgcct ggctgatacg
     3361 ttggtcctcg cgccagctta agacgctaat ccctaactgc tggcggaaaa gatgtgacag
     3421 acgcgacggc gacaagcaaa catgctgtgc gacgctggcg atatcaaaat tgctgtctgc
     3481 caggtgatcg ctgatgtact gacaagcctc gcgtacccga ttatccatcg gtggatggag
     3541 cgactcgtta atcgcttcca tgcgccgcag taacaattgc tcaagcagat ttatcgccag
     3601 cagctccgaa tagcgccctt ccccttgccc ggcgttaatg atttgcccaa acaggtcgct
     3661 gaaatgcggc tggtgcgctt catccgggcg aaagaacccc gtattggcaa atattgacgg
     3721 ccagttaagc cattcatgcc agtaggcgcg cggacgaaag taaacccact ggtgatacca
     3781 ttcgcgagcc tccggatgac gaccgtagtg atgaatctct cctggcggga acagcaaaat
     3841 atcacccggt cggcaaacaa attctcgtcc ctgatttttc accaccccct gaccgcgaat
     3901 ggtgagattg agaatataac ctttcattcc cagcggtcgg tcgataaaaa aatcgagata
     3961 accgttggcc tcaatcggcg ttaaacccgc caccagatgg gcattaaacg agtatcccgg
     4021 cagcagggga tcattttgcg cttcagccat acttttcata ctcccgccat tcagagaaga
     4081 aaccaattgt ccatattgca tcagacattg ccgtcactgc gtcttttact ggctcttctc
     4141 gctaaccaaa ccggtaaccc cgcttattaa aagcattctg taacaaagcg ggaccaaagc
     4201 catgacaaaa acgcgtaaca aaagtgtcta taatcacggc agaaaagtcc acattgatta
     4261 tttgcacggc gtcacacttt gctatgccat agcattttta tccataagat tagcggatcc
     4321 tacctgacgc tttttatcgc aactctctac tgtttctcca tacccgtttt tttgggctag
     4381 ctactagaga aagaggagaa atactagatg cgtaaaggag aagaactttt cactggagtt
     4441 gtcccaattc ttgttgaatt agatggtgat gttaatgggc acaaattttc tgtcagtgga
     4501 gagggtgaag gtgatgcaac atacggaaaa cttaccctta aatttatttg cactactgga
     4561 aaactacctg ttccatggcc aacacttgtc actactttcg gttatggtgt tcaatgcttt
     4621 gcgagatacc cagatcatat gaaacagcat gactttttca agagtgccat gcccgaaggt
     4681 tatgtacagg aaagaactat atttttcaaa gatgacggga actacaagac acgtgctgaa
     4741 gtcaagtttg aaggtgatac ccttgttaat agaatcgagt taaaaggtat tgattttaaa
     4801 gaagatggaa acattcttgg acacaaattg gaatacaact ataactcaca caatgtatac
     4861 atcatggcag acaaacaaaa gaatggaatc aaagttaact tcaaaattag acacaacatt
     4921 gaagatggaa gcgttcaact agcagaccat tatcaacaaa atactccaat tggcgatggc
     4981 cctgtccttt taccagacaa ccattacctg tccacacaat ctgccctttc gaaagatccc
     5041 aacgaaaaga gagaccacat ggtccttctt gagtttgtaa cagctgctgg gattacacat
     5101 ggcatggatg aactatacaa ataataa
//
