LOCUS       sequence-51665-e        8414 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 8414)
  AUTHORS   Andersson CR, Tsinoremas NF, Shelton J, Lebedeva NV, Yarrow J, Min 
            H, Golden SS
  TITLE     Application of bioluminescence to the study of circadian rhythms in 
            cyanobacteria.
  JOURNAL   Methods Enzymol. 2000;305:527-42.
  PUBMED    10812624
REFERENCE   2  (bases 1 to 8414)
  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: "Methods 
            Enzymol."; date: "2000"; volume: "305"; pages: "527-42"
COMMENT     Sequence Label: pAM1573
FEATURES             Location/Qualifiers
     source          1..8414
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(1569..1596)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(1688..1774)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     promoter        complement(2352..2456)
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2659..2761
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             2762..3421
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(2828..2847)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     primer_bind     complement(4485..4504)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             5966..6157
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(6151..6173)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    6259..6399
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     6314..6333
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(6414..6431)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6585..7173)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6665..6684)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7344..8135,8136..8204))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(8205..8309)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 agcttgtcat ctgccggatg aggcaaaacc ctgcctacgg cgcgattaca tcgtcccagc
       61 gcgatcgctc ttactgttga tggctcgtgc ttaaaaacaa tgcaaacttc accgtttcag
      121 ctggtgattt tcgactgtga tggtgtgctt gttgatagcg gaacgcatca ctaatcgcgt
      181 ctttgcagac atgctcaatg aactgggtct gttggtgact ttggatgaca tgtttgagca
      241 gtttgtgggt cattccatgg ctgactgtct caaactaatt gagcgacggt taggcaatcc
      301 tccaccccct gactttgttc agcactatca acgccgtacc cgtatcgcgt tagaaacgca
      361 tctacaagcc gttcctgggg ttgaagaggc tttggatgct cttgaattgc cctactgtgt
      421 tgcgtccagt ggtgatcatc aaaagatgcg aaccacactg agcctgacga agctctggcc
      481 acgatttgag ggacgaatct tcagcgtgac tgaagtacct cgcggcaagc catttcccga
      541 tgtctttttg ttggccgccg atcgcttcgg ggttaatcct acggcctgcg ctgtgatcga
      601 agacaccccc ttgggagtag cggcaggcgt ggcggcagga atgcaagtgt ttggctacgc
      661 gggttccatg cccgcttggc gtctgcaaga agccggtgcc catctcattt ttgacgatat
      721 gcgactgctg cccagtctgc tccaatcgtc gccaaaagat aactccacag cattgcccaa
      781 tccctaaccc ctgctcgcgc cgcaactaca cactaaaccg ttcctgcgcg atcgctctta
      841 ctgttgatgg ctcgtgctta aaaacaatgc aaccctaacc gtttcagctg gtgattttcg
      901 gacgatttgg cttacaggga taactgagag tcaacagcct ctgtccgtca ttgcacaccc
      961 atccatgcac tggggacttg actcatgctg aatcacattt cccttgtcca ttgggcgaga
     1021 ggggagggga atcttctgga ctcttcacta agcggcgatc gcaggttctt ctacccaagc
     1081 agtggcgatc gcttgattgc agtcttcaat gctggcctct gcagccatcg ccgccaccaa
     1141 agcatcgtag gcgggacgtt gttgctccag taaagtcttc gcccgtaaca atccccagcg
     1201 actgcgtaaa tccgcttcgg caggattgcg atcgagttgc cgccacagtt gtttccactg
     1261 ggcgcgatcg tcagctcccc cttccacgtt gccgtagacc agttgctctg ccgctgcacc
     1321 ggccatcaac acctgacacc actgttccag cgatcgctga ctgagttgcc cctgtgcggc
     1381 ttcggcttct agcgcagctg cttggaactg cacacccccg cgaccaggtt gtccttggcg
     1441 cagcgcttcc cacgctgaga gggtgtagcc cgtcacgggt aaccgatatc gtcgacaggc
     1501 ctctagaccc gggctcgagc tagcaagctt ggccggatcc ggccggatcc ggagtttgta
     1561 gaaacgcaaa aaggccatcc gtcaggatgg ccttctgctt aatttgatgc ctggcagttt
     1621 atggcgggcg tcctgcccgc caccctccgg gccgttgctt cgcaacgttc aaatccgctc
     1681 ccggcggatt tgtcctactc aggagagcgt tcaccgacaa acaacagata aaacgaaagg
     1741 cccagtcttt cgactgagcc tttcgtttta tttgatgcct ggcagttccc tactctcgca
     1801 tggggagacc ccacactacc atcggcgcta cggcgtttca cttctgagtt cggcatgggg
     1861 tcaggtggga ccaccgcgct actgccgcca ggcaaattct gttttatcag ccgttacccc
     1921 acctactagc taatcccatc tgggcacatc cgatggcaag aggcccgaag gtccccctct
     1981 ttggtcttgc gacgttatgc ggtattagct accgtttcca gtagttatcc ccctccatca
     2041 ggcagtttcc cagacattac tcacccgtcc gccactcgtc agcaaagaag caagcttaga
     2101 tccgtcacac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     2161 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     2221 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     2281 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     2341 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     2401 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     2461 tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat
     2521 aaaaataggc gtatcacgag gccctttcgt cttcgaataa atacctgtga cggaagatca
     2581 cttcgcagaa taaataaatc ctggtgtccc tgttgatacc gggaagccct gggccaactt
     2641 ttggcgaaaa tgagacgttg atcggcacgt aagaggttcc aactttcacc ataatgaaat
     2701 aagatcacta ccgggcgtat tttttgagtt atcgagattt tcaggagcta aggaagctaa
     2761 aatggagaaa aaaatcactg gatataccac cgttgatata tcccaatggc atcgtaaaga
     2821 acattttgag gcatttcagt cagttgctca atgtacctat aaccagaccg ttcagctgga
     2881 tattacggcc tttttaaaga ccgtaaagaa aaataagcac aagttttatc cggcctttat
     2941 tcacattctt gcccgcctga tgaatgctca tccggaattc cgtatggcaa tgaaagacgg
     3001 tgagctggtg atatgggata gtgttcaccc ttgttacacc gttttccatg agcaaactga
     3061 aacgttttca tcgctctgga gtgaatacca cgacgatttc cggcagtttc tacacatata
     3121 ttcgcaagat gtggcgtgtt acggtgaaaa cctggcctat ttccctaaag ggtttattga
     3181 gaatatgttt ttcgtctcag ccaatccctg ggtgagtttc accagttttg atttaaacgt
     3241 ggccaatatg gacaacttct tcgcccccgt tttcaccatg ggcaaatatt atacgcaagg
     3301 cgacaaggtg ctgatgccgc tggcgattca ggttcatcat gccgtttgtg atggcttcca
     3361 tgtcggcaga atgcttaatg aattacaaca gtactgcgat gagtggcagg gcggggcgta
     3421 atttttttaa ggcagttatt ggtgccctta aacgcctggt tgctacgcct gaataagtga
     3481 taataagcgg atgaatggca gaaattcgaa agcaaattcg acccggtcgt cggttcaggg
     3541 cagggtcgtt aaatagccgc ttatgtctat tgctggttta ccggtttatt gactaccgga
     3601 agcagtgtga ccgtgtgctt ctcaaatgcc tgaggccagt ttgctcaggc tctccccgtg
     3661 gaggtaataa ttgacgatat gatcgacgga tctggtaacc ccagcgcggt tgctaccaag
     3721 tagtgacccg cttcgtgatg caaaatccgc tgacgatatt cgggcgatcg ctgctgaatg
     3781 ccatcgagca gtaacgtggc accccgcccc tgccaagtca ccgcatccag actgaacagc
     3841 accaagaggc taaaacccaa tcccgccggt agcagcggag aactacccag cattggtccc
     3901 accaaagcta atgccgtcgt ggtaaaaatc gcgatcgccg tcagactcaa gcccagttcg
     3961 ctcatgcttc ctcatctagg tcacagtctt cggcgatcgc atcgatctga tgctgcagca
     4021 agcgttttcc ataccggcga tcgcgccgtc gccctttcgc tgccgtggcc cgcttacgag
     4081 ctcgtttatc gaccacgatc gcatccaaat ccgcgatcgc ttcccagtcc ggcaattcag
     4141 tctggggcgt ccgtttcatt aatcctgatc aggcacgaaa ttgctgtgcg tagtatcgcg
     4201 catagcggcc agcctctgcc aacagcgcat cgtgattgcc tgcctcaaca atctggccgc
     4261 gctccatcac caagatgcgg ctggcattac gaaccgtagc cagacggtga gcaatgataa
     4321 agaccgtccg tccctgcatc acccgttcta gggcctcttg caccaaggtt tcggactcgg
     4381 aatcaagcgc cgaagtcgcc tcatccagaa ttaaaatgcg tggatcctct acgccggacg
     4441 catcgtggcc ggcatcaccg gcgccacagg tgcggttgct ggcgcctata tcgccgacat
     4501 caccgatggg gaagatcggg ctcgccactt cgggctcatg agcgcttgtt tcggcgtggg
     4561 tatggtggca ggccccgtgg ccgggggact gttgggcgcc atctccttgc atgcaccatt
     4621 ccttgcggcg gcggtgctca acggcctcaa cctactactg ggctgcttcc taatgcagga
     4681 gtcgcataag ggagagcgtc gatcgaccga tgcccttgag agccttcaac ccagtcagct
     4741 ccttccggtg ggcgcggggc atgactatcg tcgccgcact tatgactgtc ttctttatca
     4801 tgcaactcgt aggacaggtg ccggcagcgc tctgggtcat tttcggcgag gaccgctttc
     4861 gctggagcgc gacgatgatc ggcctgtcgc ttgcggtatt cggaatcttg cacgccctcg
     4921 ctcaagcctt cgtcactggt cccgccacca aacgtttcgg cgagaagcag gccattatcg
     4981 ccggcatggc ggccgacgcg ctgggctacg tcttgctggc gttcgcgacg cgaggctgga
     5041 tggccttccc cattatgatt cttctcgctt ccggcggcat cgggatgccc gcgttgcagg
     5101 ccatgctgtc caggcaggta gatgacgacc atcagggaca gcttcaagga tcgctcgcgg
     5161 ctcttaccag cctaacttcg atcactggac cgctgatcgt cacggcgatt tatgccgcct
     5221 cggcgagcac atggaacggg ttggcatgga ttgtaggcgc cgccctatac cttgtctgcc
     5281 tccccgcgtt gcgtcgcggt gcatggagcc gggccacctc gacctgaatg gaagccggcg
     5341 gcacctcgct aacggattca ccactccaag aattggagcc aatcaattct tgcggagaac
     5401 tgtgaatgcg caaaccaacc cttggcagaa catatccatc gcgtccgcca tctccagcag
     5461 ccgcacgcgg cgcatctcgg gcagcgttgg gtcctggcca cgggtgcgca tgatcgtgct
     5521 cctgtcgttg aggacccggc taggctggcg gggttgcctt actggttagc agaatgaatc
     5581 accgatacgc gagcgaacgt gaagcgactg ctgctgcaaa acgtctgcga cctgagcaac
     5641 aacatgaatg gtcttcggtt tccgtgtttc gtaaagtctg gaaacgcgga agtcagcgcc
     5701 ctgcaccatt atgttccgga tctgcatcgc aggatgctgc tggctaccct gtggaacacc
     5761 tacatctgta ttaacgaagc gctggcattg accctgagtg atttttctct ggtcccgccg
     5821 catccatacc gccagttgtt taccctcaca acgttccagt aaccgggcat gttcatcatc
     5881 agtaacccgt atcgtgagca tcctctctcg tttcatcggt atcattaccc ccatgaacag
     5941 aaatccccct tacacggagg catcagtgac caaacaggaa aaaaccgccc ttaacatggc
     6001 ccgctttatc agaagccaga cattaacgct tctggagaaa ctcaacgagc tggacgcgga
     6061 tgaacaggca gacatctgtg aatcgcttca cgaccacgct gatgagcttt accgcagctg
     6121 cctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt
     6181 cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg
     6241 tgttggcggg tgtcggggcg cagccatgac ccagtcacgt agcgatagcg gagtgtatac
     6301 tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccatat gcggtgtgaa
     6361 ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc gctcttccgc ttcctcgctc
     6421 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     6481 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     6541 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     6601 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     6661 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     6721 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     6781 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     6841 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     6901 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     6961 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     7021 agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     7081 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     7141 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     7201 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     7261 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     7321 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     7381 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     7441 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     7501 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     7561 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     7621 tcgccagtta atagtttgcg caacgttgtt gccattgctg caggcatcgt ggtgtcacgc
     7681 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     7741 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     7801 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     7861 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     7921 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa cacgggataa taccgcgcca
     7981 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     8041 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     8101 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     8161 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     8221 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     8281 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
     8341 taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt
     8401 cgtcttcaag aatt
//
