LOCUS       sequence-64701-e        5091 bp    DNA     circular SYN 15-APR-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5091)
  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 5091)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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-ECK120010800
FEATURES             Location/Qualifiers
     source          1..5091
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             71..82
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             89..766
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     terminator      811..882
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      898..925
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     primer_bind     complement(1019..1036)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1190..1778)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1270..1289)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1949..2740,2741..2809))
                     /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     2572..2591
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2810..2914)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2982..3000
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(3004..3047)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     CDS             complement(3136..4014)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        4041..4325
                     /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     4248..4267
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     RBS             4354..4365
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             4372..5085
                     /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(4402..4430)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     5025..5045
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
ORIGIN
        1 gaattcagtt tgttcgcccg gtagttgtga cgctaccggg ttcttttcga actagtagcg
       61 gccgctgcag aaagaggaga aatactagat ggcttcctcc gaagacgtta tcaaagagtt
      121 catgcgtttc aaagttcgta tggaaggttc cgttaacggt cacgagttcg aaatcgaagg
      181 tgaaggtgaa ggtcgtccgt acgaaggtac ccagaccgct aaactgaaag ttaccaaagg
      241 tggtccgctg ccgttcgctt gggacatcct gtccccgcag ttccagtacg gttccaaagc
      301 ttacgttaaa cacccggctg acatcccgga ctacctgaaa ctgtccttcc cggaaggttt
      361 caaatgggaa cgtgttatga acttcgaaga cggtggtgtt gttaccgtta cccaggactc
      421 ctccctgcaa gacggtgagt tcatctacaa agttaaactg cgtggtacca acttcccgtc
      481 cgacggtccg gttatgcaga aaaaaaccat gggttgggaa gcttccaccg aacgtatgta
      541 cccggaagac ggtgctctga aaggtgaaat caaaatgcgt ctgaaactga aagacggtgg
      601 tcactacgac gctgaagtta aaaccaccta catggctaaa aaaccggttc agctgccggg
      661 tgcttacaaa accgacatca aactggacat cacctcccac aacgaagact acaccatcgt
      721 tgaacagtac gaacgtgctg aaggtcgtca ctccaccggt gcttaataac gctgatagtg
      781 ctagtgtaga tcgctactag agccaggcat caaataaaac gaaaggctca gtcgaaagac
      841 tgggcctttc gttttatctg ttgtttgtcg gtgaacgctc tctactagag tcacactggc
      901 tcaccttcgg gtgggccttt ctgcgtttat atactagagt caattgaggt agaggtacac
      961 acgcgaactc cgatagccaa ttcagagtaa taaactgtga taatcaatgc agtgcttcct
     1021 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     1081 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     1141 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     1201 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     1261 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     1321 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     1381 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     1441 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     1501 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     1561 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     1621 acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     1681 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     1741 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     1801 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     1861 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     1921 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     1981 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     2041 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     2101 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     2161 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     2221 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     2281 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     2341 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     2401 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     2461 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     2521 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     2581 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     2641 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     2701 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     2761 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     2821 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     2881 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     2941 acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc
     3001 agaatttcag ataaaaaaaa tccttagctt tcgctaagga tgatttctgg aattgaattg
     3061 aggtagaggt acacacgcga actccgatag ccaattcaga gtaataaact gtgataatca
     3121 atgcatgtac tagagttatg acaacttgac ggctacatca ttcacttttt cttcacaacc
     3181 ggcacggaac tcgctcgggc tggccccggt gcatttttta aatacccgcg agaaatagag
     3241 ttgatcgtca aaaccaacat tgcgaccgac ggtggcgata ggcatccggg tggtgctcaa
     3301 aagcagcttc gcctggctga tacgttggtc ctcgcgccag cttaagacgc taatccctaa
     3361 ctgctggcgg aaaagatgtg acagacgcga cggcgacaag caaacatgct gtgcgacgct
     3421 ggcgatatca aaattgctgt ctgccaggtg atcgctgatg tactgacaag cctcgcgtac
     3481 ccgattatcc atcggtggat ggagcgactc gttaatcgct tccatgcgcc gcagtaacaa
     3541 ttgctcaagc agatttatcg ccagcagctc cgaatagcgc ccttcccctt gcccggcgtt
     3601 aatgatttgc ccaaacaggt cgctgaaatg cggctggtgc gcttcatccg ggcgaaagaa
     3661 ccccgtattg gcaaatattg acggccagtt aagccattca tgccagtagg cgcgcggacg
     3721 aaagtaaacc cactggtgat accattcgcg agcctccgga tgacgaccgt agtgatgaat
     3781 ctctcctggc gggaacagca aaatatcacc cggtcggcaa acaaattctc gtccctgatt
     3841 tttcaccacc ccctgaccgc gaatggtgag attgagaata taacctttca ttcccagcgg
     3901 tcggtcgata aaaaaatcga gataaccgtt ggcctcaatc ggcgttaaac ccgccaccag
     3961 atgggcatta aacgagtatc ccggcagcag gggatcattt tgcgcttcag ccatactttt
     4021 catactcccg ccattcagag aagaaaccaa ttgtccatat tgcatcagac attgccgtca
     4081 ctgcgtcttt tactggctct tctcgctaac caaaccggta accccgctta ttaaaagcat
     4141 tctgtaacaa agcgggacca aagccatgac aaaaacgcgt aacaaaagtg tctataatca
     4201 cggcagaaaa gtccacattg attatttgca cggcgtcaca ctttgctatg ccatagcatt
     4261 tttatccata agattagcgg atcctacctg acgcttttta tcgcaactct ctactgtttc
     4321 tccatacccg tttttttggg ctagctacta gagaaagagg agaaatacta gatgcgtaaa
     4381 ggagaagaac ttttcactgg agttgtccca attcttgttg aattagatgg tgatgttaat
     4441 gggcacaaat tttctgtcag tggagagggt gaaggtgatg caacatacgg aaaacttacc
     4501 cttaaattta tttgcactac tggaaaacta cctgttccat ggccaacact tgtcactact
     4561 ttcggttatg gtgttcaatg ctttgcgaga tacccagatc atatgaaaca gcatgacttt
     4621 ttcaagagtg ccatgcccga aggttatgta caggaaagaa ctatattttt caaagatgac
     4681 gggaactaca agacacgtgc tgaagtcaag tttgaaggtg atacccttgt taatagaatc
     4741 gagttaaaag gtattgattt taaagaagat ggaaacattc ttggacacaa attggaatac
     4801 aactataact cacacaatgt atacatcatg gcagacaaac aaaagaatgg aatcaaagtt
     4861 aacttcaaaa ttagacacaa cattgaagat ggaagcgttc aactagcaga ccattatcaa
     4921 caaaatactc caattggcga tggccctgtc cttttaccag acaaccatta cctgtccaca
     4981 caatctgccc tttcgaaaga tcccaacgaa aagagagacc acatggtcct tcttgagttt
     5041 gtaacagctg ctgggattac acatggcatg gatgaactat acaaataata a
//
