LOCUS       sequence-51666-e        8079 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 8079)
  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 8079)
  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: pAM1579
FEATURES             Location/Qualifiers
     source          1..8079
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(1568..1595)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(1687..1773)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(2221..2240)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2232..3047
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             2232..3047
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(2305..2324)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     complement(4150..4169)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             5631..5822
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(5816..5838)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    5924..6064
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     5979..5998
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(6079..6096)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6250..6838)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6330..6349)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7009..7800,7801..7869))
                     /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"
     primer_bind     7632..7651
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7870..7974)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     8042..8060
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 agcttgtcat ctgccggatg aggcaaaacc ctgcctacgg cgcgattaca tcgtcccagc
       61 gcgatcgctc ttactgttga tggctcgtgc ttaaaaacaa tgcaaacttc accgtttcag
      121 ctggtgattt tcgactgtga tggtgtgctt gttgatagcg aacgcatcac taatcgcgtc
      181 tttgcagaca tgctcaatga actgggtctg ttggtgactt tggatgacat gtttgagcag
      241 tttgtgggtc attccatggc tgactgtctc aaactaattg agcgacggtt aggcaatcct
      301 ccaccccctg actttgttca gcactatcaa cgccgtaccc gtatcgcgtt agaaacgcat
      361 ctacaagccg ttcctggggt tgaagaggct ttggatgctc ttgaattgcc ctactgtgtt
      421 gcgtccagtg gtgatcatca aaagatgcga accacactga gcctgacgaa gctctggcca
      481 cgatttgagg gacgaatctt cagcgtgact gaagtacctc gcggcaagcc atttcccgat
      541 gtctttttgt tggccgccga tcgcttcggg gttaatccta cggcctgcgc tgtgatcgaa
      601 gacaccccct tgggagtagc ggcaggcgtg gcggcaggaa tgcaagtgtt tggctacgcg
      661 ggttccatgc ccgcttggcg tctgcaagaa gccggtgccc atctcatttt tgacgatatg
      721 cgactgctgc ccagtctgct ccaatcgtcg ccaaaagata actccacagc attgcccaat
      781 ccctaacccc tgctcgcgcc gcaactacac actaaaccgt tcctgcgcga tcgctcttac
      841 tgttgatggc tcgtgcttaa aaacaatgca accctaaccg tttcagctgg tgattttcgg
      901 acgatttggc ttacagggat aactgagagt caacagcctc tgtccgtcat tgcacaccca
      961 tccatgcact ggggacttga ctcatgctga atcacatttc ccttgtccat tgggcgagag
     1021 gggaggggaa tcttctggac tcttcactaa gcggcgatcg caggttcttc tacccaagca
     1081 gtggcgatcg cttgattgca gtcttcaatg ctggcctctg cagccatcgc cgccaccaaa
     1141 gcatcgtagg cgggacgttg ttgctccagt aaagtcttcg cccgtaacaa tccccagcga
     1201 ctgcgtaaat ccgcttcggc aggattgcga tcgagttgcc gccacagttg tttccactgg
     1261 gcgcgatcgt cagctccccc ttccacgttg ccgtagacca gttgctctgc cgctgcaccg
     1321 gccatcaaca cctgacacca ctgttccagc gatcgctgac tgagttgccc ctgtgcggct
     1381 tcggcttcta gcgcagctgc ttggaactgc acacccccgc gaccaggttg tccttggcgc
     1441 agcgcttccc acgctgagag ggtgtagccc gtcacgggta accgatatcg tcgacaggcc
     1501 tctagacccg ggctcgagct agcaagcttg gccggatccg gccggatccg gagtttgtag
     1561 aaacgcaaaa aggccatccg tcaggatggc cttctgctta atttgatgcc tggcagttta
     1621 tggcgggcgt cctgcccgcc accctccggg ccgttgcttc gcaacgttca aatccgctcc
     1681 cggcggattt gtcctactca ggagagcgtt caccgacaaa caacagataa aacgaaaggc
     1741 ccagtctttc gactgagcct ttcgttttat ttgatgcctg gcagttccct actctcgcat
     1801 ggggagaccc cacactacca tcggcgctac ggcgtttcac ttctgagttc ggcatggggt
     1861 caggtgggac caccgcgcta ctgccgccag gcaaattctg ttttatcagc cgttacccca
     1921 cctactagct aatcccatct gggcacatcc gatggcaaga ggcccgaagg tccccctctt
     1981 tggtcttgcg acgttatgcg gtattagcta ccgtttccag tagttatccc cctccatcag
     2041 gcagtttccc agacattact cacccgtccg ccactcgtca gcaaagaagc aagcttagat
     2101 cgacctgcag gggggggggg gaaagccacg ttgtgtctca aaatctctga tgttacattg
     2161 cacaagataa aaatatatca tcatgaacaa taaaactgtc tgcttacata aacagtaata
     2221 caaggggtgt tatgagccat attcaacggg aaacgtcttg ctcgaggccg cgattaaatt
     2281 ccaacatgga tgctgattta tatgggtata aatgggctcg cgataatgtc gggcaatcag
     2341 gtgcgacaat ctatcgattg tatgggaagc ccgatgcgcc agagttgttt ctgaaacatg
     2401 gcaaaggtag cgttgccaat gatgttacag atgagatggt cagactaaac tggctgacgg
     2461 aatttatgcc tcttccgacc atcaagcatt ttatccgtac tcctgatgat gcatggttac
     2521 tcaccactgc gatccccggg aaaacagcat tccaggtatt agaagaatat cctgattcag
     2581 gtgaaaatat tgttgatgcg ctggcagtgt tcctgcgccg gttgcattcg attcctgttt
     2641 gtaattgtcc ttttaacagc gatcgcgtat ttcgtctcgc tcaggcgcaa tcacgaatga
     2701 ataacggttt ggttgatgcg agtgattttg atgacgagcg taatggctgg cctgttgaac
     2761 aagtctggaa agaaatgcat aagcttttgc cattctcacc ggattcagtc gtcactcatg
     2821 gtgatttctc acttgataac cttatttttg acgaggggaa attaataggt tgtattgatg
     2881 ttggacgagt cggaatcgca gaccgatacc aggatcttgc catcctatgg aactgcctcg
     2941 gtgagttttc tccttcatta cagaaacggc tttttcaaaa atatggtatt gataatcctg
     3001 atatgaataa attgcagttt catttgatgc tcgatgagtt tttctaatca gaattggtta
     3061 attggttgta acactggcag agcattacgc tgacttgacg ggacggcggc tttgttgaat
     3121 aaatcgaact tttgctgagt tgaaggatca gatcacgcat cttcccgaca acgcagaccg
     3181 ttccgtggca aagcaaaagt tcaaaatcac caactggtcc acctacaaca aagctctcat
     3241 caaccgtggc tccctcactt tctggctgga tgatggggcg attcaggcct ggtatgagtc
     3301 agcaacacct tcttcacgag gcagacctca gcgccccccc ccccctgcag gtcgatctgg
     3361 taaccccagc gcggttgcta ccaagtagtg acccgcttcg tgatgcaaaa tccgctgacg
     3421 atattcgggc gatcgctgct gaatgccatc gagcagtaac gtggcacccc gcccctgcca
     3481 agtcaccgca tccagactga acagcaccaa gaggctaaaa cccaatcccg ccggtagcag
     3541 cggagaacta cccagcattg gtcccaccaa agctaatgcc gtcgtggtaa aaatcgcgat
     3601 cgccgtcaga ctcaagccca gttcgctcat gcttcctcat ctaggtcaca gtcttcggcg
     3661 atcgcatcga tctgatgctg cagcaagcgt tttccatacc ggcgatcgcg ccgtcgccct
     3721 ttcgctgccg tggcccgctt acgagctcgt ttatcgacca cgatcgcatc caaatccgcg
     3781 atcgcttccc agtccggcaa ttcagtctgg ggcgtccgtt tcattaatcc tgatcaggca
     3841 cgaaattgct gtgcgtagta tcgcgcatag cggccagcct ctgccaacag cgcatcgtga
     3901 ttgcctgcct caacaatctg gccgcgctcc atcaccaaga tgcggctggc attacgaacc
     3961 gtagccagac ggtgagcaat gataaagacc gtccgtccct gcatcacccg ttctagggcc
     4021 tcttgcacca aggtttcgga ctcggaatca agcgccgaag tcgcctcatc cagaattaaa
     4081 atgcgtggat cctctacgcc ggacgcatcg tggccggcat caccggcgcc acaggtgcgg
     4141 ttgctggcgc ctatatcgcc gacatcaccg atggggaaga tcgggctcgc cacttcgggc
     4201 tcatgagcgc ttgtttcggc gtgggtatgg tggcaggccc cgtggccggg ggactgttgg
     4261 gcgccatctc cttgcatgca ccattccttg cggcggcggt gctcaacggc ctcaacctac
     4321 tactgggctg cttcctaatg caggagtcgc ataagggaga gcgtcgatcg accgatgccc
     4381 ttgagagcct tcaacccagt cagctccttc cggtgggcgc ggggcatgac tatcgtcgcc
     4441 gcacttatga ctgtcttctt tatcatgcaa ctcgtaggac aggtgccggc agcgctctgg
     4501 gtcattttcg gcgaggaccg ctttcgctgg agcgcgacga tgatcggcct gtcgcttgcg
     4561 gtattcggaa tcttgcacgc cctcgctcaa gccttcgtca ctggtcccgc caccaaacgt
     4621 ttcggcgaga agcaggccat tatcgccggc atggcggccg acgcgctggg ctacgtcttg
     4681 ctggcgttcg cgacgcgagg ctggatggcc ttccccatta tgattcttct cgcttccggc
     4741 ggcatcggga tgcccgcgtt gcaggccatg ctgtccaggc aggtagatga cgaccatcag
     4801 ggacagcttc aaggatcgct cgcggctctt accagcctaa cttcgatcac tggaccgctg
     4861 atcgtcacgg cgatttatgc cgcctcggcg agcacatgga acgggttggc atggattgta
     4921 ggcgccgccc tataccttgt ctgcctcccc gcgttgcgtc gcggtgcatg gagccgggcc
     4981 acctcgacct gaatggaagc cggcggcacc tcgctaacgg attcaccact ccaagaattg
     5041 gagccaatca attcttgcgg agaactgtga atgcgcaaac caacccttgg cagaacatat
     5101 ccatcgcgtc cgccatctcc agcagccgca cgcggcgcat ctcgggcagc gttgggtcct
     5161 ggccacgggt gcgcatgatc gtgctcctgt cgttgaggac ccggctaggc tggcggggtt
     5221 gccttactgg ttagcagaat gaatcaccga tacgcgagcg aacgtgaagc gactgctgct
     5281 gcaaaacgtc tgcgacctga gcaacaacat gaatggtctt cggtttccgt gtttcgtaaa
     5341 gtctggaaac gcggaagtca gcgccctgca ccattatgtt ccggatctgc atcgcaggat
     5401 gctgctggct accctgtgga acacctacat ctgtattaac gaagcgctgg cattgaccct
     5461 gagtgatttt tctctggtcc cgccgcatcc ataccgccag ttgtttaccc tcacaacgtt
     5521 ccagtaaccg ggcatgttca tcatcagtaa cccgtatcgt gagcatcctc tctcgtttca
     5581 tcggtatcat tacccccatg aacagaaatc ccccttacac ggaggcatca gtgaccaaac
     5641 aggaaaaaac cgcccttaac atggcccgct ttatcagaag ccagacatta acgcttctgg
     5701 agaaactcaa cgagctggac gcggatgaac aggcagacat ctgtgaatcg cttcacgacc
     5761 acgctgatga gctttaccgc agctgcctcg cgcgtttcgg tgatgacggt gaaaacctct
     5821 gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac
     5881 aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggcgcagcc atgacccagt
     5941 cacgtagcga tagcggagtg tatactggct taactatgcg gcatcagagc agattgtact
     6001 gagagtgcac catatgcggt gtgaaatacc gcacagatgc gtaaggagaa aataccgcat
     6061 caggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     6121 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     6181 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     6241 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     6301 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     6361 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     6421 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     6481 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     6541 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     6601 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     6661 gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa
     6721 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     6781 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     6841 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     6901 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     6961 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     7021 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     7081 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     7141 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     7201 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     7261 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     7321 tgctgcaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     7381 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     7441 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     7501 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     7561 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     7621 gtcaacacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     7681 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     7741 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     7801 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     7861 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     7921 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     7981 tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa
     8041 aaataggcgt atcacgaggc cctttcgtct tcaagaatt
//
