LOCUS       sequence-64688-e        5108 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 5108)
  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 5108)
  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-L3S2P21
FEATURES             Location/Qualifiers
     source          1..5108
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             88..99
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             106..783
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     terminator      828..899
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      915..942
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     primer_bind     complement(1036..1053)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1207..1795)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1287..1306)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1966..2757,2758..2826))
                     /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     2589..2608
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2827..2931)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2999..3017
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(3021..3064)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     CDS             complement(3153..4031)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        4058..4342
                     /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     4265..4284
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     RBS             4371..4382
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             4389..5102
                     /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(4419..4447)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     5042..5062
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
ORIGIN
        1 gaattcctcg gtaccaaatt ccagaaaaga ggcctcccga aaggggggcc ttttttcgtt
       61 ttggtccact agtagcggcc gctgcagaaa gaggagaaat actagatggc ttcctccgaa
      121 gacgttatca aagagttcat gcgtttcaaa gttcgtatgg aaggttccgt taacggtcac
      181 gagttcgaaa tcgaaggtga aggtgaaggt cgtccgtacg aaggtaccca gaccgctaaa
      241 ctgaaagtta ccaaaggtgg tccgctgccg ttcgcttggg acatcctgtc cccgcagttc
      301 cagtacggtt ccaaagctta cgttaaacac ccggctgaca tcccggacta cctgaaactg
      361 tccttcccgg aaggtttcaa atgggaacgt gttatgaact tcgaagacgg tggtgttgtt
      421 accgttaccc aggactcctc cctgcaagac ggtgagttca tctacaaagt taaactgcgt
      481 ggtaccaact tcccgtccga cggtccggtt atgcagaaaa aaaccatggg ttgggaagct
      541 tccaccgaac gtatgtaccc ggaagacggt gctctgaaag gtgaaatcaa aatgcgtctg
      601 aaactgaaag acggtggtca ctacgacgct gaagttaaaa ccacctacat ggctaaaaaa
      661 ccggttcagc tgccgggtgc ttacaaaacc gacatcaaac tggacatcac ctcccacaac
      721 gaagactaca ccatcgttga acagtacgaa cgtgctgaag gtcgtcactc caccggtgct
      781 taataacgct gatagtgcta gtgtagatcg ctactagagc caggcatcaa ataaaacgaa
      841 aggctcagtc gaaagactgg gcctttcgtt ttatctgttg tttgtcggtg aacgctctct
      901 actagagtca cactggctca ccttcgggtg ggcctttctg cgtttatata ctagagtcaa
      961 ttgaggtaga ggtacacacg cgaactccga tagccaattc agagtaataa actgtgataa
     1021 tcaatgcagt gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     1081 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     1141 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     1201 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     1261 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     1321 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     1381 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     1441 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     1501 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     1561 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     1621 gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc
     1681 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     1741 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     1801 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     1861 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     1921 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     1981 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     2041 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     2101 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     2161 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     2221 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     2281 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     2341 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     2401 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     2461 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     2521 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     2581 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     2641 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     2701 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     2761 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     2821 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     2881 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     2941 ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa
     3001 taggcgtatc acgaggcaga atttcagata aaaaaaatcc ttagctttcg ctaaggatga
     3061 tttctggaat tgaattgagg tagaggtaca cacgcgaact ccgatagcca attcagagta
     3121 ataaactgtg ataatcaatg catgtactag agttatgaca acttgacggc tacatcattc
     3181 actttttctt cacaaccggc acggaactcg ctcgggctgg ccccggtgca ttttttaaat
     3241 acccgcgaga aatagagttg atcgtcaaaa ccaacattgc gaccgacggt ggcgataggc
     3301 atccgggtgg tgctcaaaag cagcttcgcc tggctgatac gttggtcctc gcgccagctt
     3361 aagacgctaa tccctaactg ctggcggaaa agatgtgaca gacgcgacgg cgacaagcaa
     3421 acatgctgtg cgacgctggc gatatcaaaa ttgctgtctg ccaggtgatc gctgatgtac
     3481 tgacaagcct cgcgtacccg attatccatc ggtggatgga gcgactcgtt aatcgcttcc
     3541 atgcgccgca gtaacaattg ctcaagcaga tttatcgcca gcagctccga atagcgccct
     3601 tccccttgcc cggcgttaat gatttgccca aacaggtcgc tgaaatgcgg ctggtgcgct
     3661 tcatccgggc gaaagaaccc cgtattggca aatattgacg gccagttaag ccattcatgc
     3721 cagtaggcgc gcggacgaaa gtaaacccac tggtgatacc attcgcgagc ctccggatga
     3781 cgaccgtagt gatgaatctc tcctggcggg aacagcaaaa tatcacccgg tcggcaaaca
     3841 aattctcgtc cctgattttt caccaccccc tgaccgcgaa tggtgagatt gagaatataa
     3901 cctttcattc ccagcggtcg gtcgataaaa aaatcgagat aaccgttggc ctcaatcggc
     3961 gttaaacccg ccaccagatg ggcattaaac gagtatcccg gcagcagggg atcattttgc
     4021 gcttcagcca tacttttcat actcccgcca ttcagagaag aaaccaattg tccatattgc
     4081 atcagacatt gccgtcactg cgtcttttac tggctcttct cgctaaccaa accggtaacc
     4141 ccgcttatta aaagcattct gtaacaaagc gggaccaaag ccatgacaaa aacgcgtaac
     4201 aaaagtgtct ataatcacgg cagaaaagtc cacattgatt atttgcacgg cgtcacactt
     4261 tgctatgcca tagcattttt atccataaga ttagcggatc ctacctgacg ctttttatcg
     4321 caactctcta ctgtttctcc atacccgttt ttttgggcta gctactagag aaagaggaga
     4381 aatactagat gcgtaaagga gaagaacttt tcactggagt tgtcccaatt cttgttgaat
     4441 tagatggtga tgttaatggg cacaaatttt ctgtcagtgg agagggtgaa ggtgatgcaa
     4501 catacggaaa acttaccctt aaatttattt gcactactgg aaaactacct gttccatggc
     4561 caacacttgt cactactttc ggttatggtg ttcaatgctt tgcgagatac ccagatcata
     4621 tgaaacagca tgactttttc aagagtgcca tgcccgaagg ttatgtacag gaaagaacta
     4681 tatttttcaa agatgacggg aactacaaga cacgtgctga agtcaagttt gaaggtgata
     4741 cccttgttaa tagaatcgag ttaaaaggta ttgattttaa agaagatgga aacattcttg
     4801 gacacaaatt ggaatacaac tataactcac acaatgtata catcatggca gacaaacaaa
     4861 agaatggaat caaagttaac ttcaaaatta gacacaacat tgaagatgga agcgttcaac
     4921 tagcagacca ttatcaacaa aatactccaa ttggcgatgg ccctgtcctt ttaccagaca
     4981 accattacct gtccacacaa tctgcccttt cgaaagatcc caacgaaaag agagaccaca
     5041 tggtccttct tgagtttgta acagctgctg ggattacaca tggcatggat gaactataca
     5101 aataataa
//
