LOCUS       sequence-180106-        6589 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  PkaiB-kaiC-ecfp expressed from NSII-Gm vector constructed by 
            seamless assembly. Fusion expressed as full length fusion protein at
            WT levels but is 5hr long period. Used only for co-localization exp.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6589)
  AUTHORS   Cohen SE, Erb ML, Selimkhanov J, Dong G, Hasty J, Pogliano J, Golden
            SS
  TITLE     Dynamic localization of the cyanobacterial circadian clock proteins.
  JOURNAL   Curr Biol. 2014 Aug 18;24(16):1836-44. doi: 
            10.1016/j.cub.2014.07.036. Epub 2014 Aug 7.
  PUBMED    25127213
REFERENCE   2  (bases 1 to 6589)
  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"; doi: 
            "10.1016/j.cub.2014.07.036"; journalName: "Curr Biol"; date: 
            "2014-08-18- 18"; volume: "24"; issue: "16"; pages: "1836-44"
COMMENT     Sequence Label: pAM5082
FEATURES             Location/Qualifiers
     source          1..6589
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      8..596
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     497..516
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     750..767
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    782..922
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(848..867)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     CDS             complement(2022..2555)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(2744..2749,2750..2766,2767..2772))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             join(5288..5290,5291..5293,5294..6007)
                     /codon_start=1
                     /product="enhanced CFP"
                     /label=ECFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(5333..5354)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5594..5613)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5941..5962
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     terminator      6031..6058
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
ORIGIN
        1 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
       61 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
      121 ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc
      181 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
      241 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
      301 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
      361 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
      421 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
      481 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
      541 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
      601 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
      661 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata
      721 ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc
      781 gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atatggtgca
      841 ctctcagtac aatctgctct gatgccgcat agttaagcca gtatacactc cgctatcgct
      901 acgtgactgg gtcatggctg cgccccgaca cccgccaaca cccgctgacg cgccctgacg
      961 ggcttgtctg ctcccgccat ccgcttacag acaagctccg cgggccacga tttgagggac
     1021 gaatcttcag cgtgactgaa gtacctcgcg gcaagccatt tcccgatgtc tttttgttgg
     1081 ccgccgatcg cttcggggtt aatcctacgg cctgcgctgt gatcgaagac acccccttgg
     1141 gagtagcggc aggcgtggcg gcaggaatgc aagtgtttgg ctacgcgggt tccatgcccg
     1201 cttggcgtct gcaagaagcc ggtgcccatc tcatttttga cgatatgcga ctgctgccca
     1261 gtctgctcca atcgtcgcca aaagataact ccacagcatt gcccaatccc taacccctgc
     1321 tcgcgccgca actacacact aaaccgttcc tgcgcgatcg ctcttactgt tgatggctcg
     1381 tgcttaaaaa caatgcaacc ctaaccgttt cagctggtga ttttcggacg atttggctta
     1441 cagggataac tgagagtcaa cagcctctgt ccgtcattgc acacccatcc atgcactggg
     1501 gacttgactc atgctgaatc acatttccct tgtccattgg gcgagagggg aggggaatct
     1561 tctggactct tcactaagcg gcgatcgcag gttcttctac ccaagcagtg gcgatcgctt
     1621 gattgcagtc ttcaatgctg gcctctgcag ccatcgccgc caccaaagca tcgtaggcgg
     1681 gacgttgttg ctccagtaaa gtcttcgccc gtaacaatcc ccagcgactg cgtaaatccg
     1741 cttcggcagg attgcgatcg agttgccgcc acagttgttt ccactgggcg cgatcgtcag
     1801 ctcccccttc cacgttgccg tagaccagtt gctctgccgc tgcaccggcc atcaacacct
     1861 gacaccactg ttccagcgat cgctgactga gttgcccctg tgcggcttcg gcttctagcg
     1921 cagctgcttg gaactgcaca cccccgctag catcgcgcgc gcgcgcgcgg atgctagcgg
     1981 cgcgccaaaa aaaagcccgc tcattaggcg ggctggccgg cttaggtggc ggtacttggg
     2041 tcaatatcaa agtgcatcac ttcttcccgt atgcccaact ttgtatagag agccactgcg
     2101 ggatcgtcac cgtaatctgc ttgcacgtag atcacataag caccaagcgc gttggcctca
     2161 tgcttgagga gattgatgag cgcggtggca atgccctgcc tccggtgctc gccggagact
     2221 gcgagatcat agatatagat ctcactacgc ggctgctcaa acttgggcag aacgtaagcc
     2281 gcgagagcgc caacaaccgc ttcttggtcg aaggcagcaa gcgcgatgaa tgtcttacta
     2341 cggagcaagt tcccgaggta atcggagtcc ggctgatgtt gggagtaggt ggctacgtct
     2401 ccgaactcac gaccgaaaag atcaagagca gcccgcatgg atttgacttg gtcagggccg
     2461 agcctacatg tgcgaatgat gcccatactt gagccaccta actttgtttt agggcgactg
     2521 ccctgctgcg taacatcgtt gctgctgcgt aacatcgttg ctgctccata acatcaaaca
     2581 tcgacccacg gcgtaacgcg cttgctgctt ggatgcccga ggcatagact gtacaaaaaa
     2641 acagtcataa caagccatga aaaccgccac tgcgccgtta ccaccgctgc gttcggtcaa
     2701 ggttctggac cagttgcgtg agcgcatacg ctacttgcat tacagtttac gaaccgaaca
     2761 ggcttatgtc aactgggttc gtgccttcat ccgtttccac ggtgtgcgtc acccggcaac
     2821 cttgggcagc agcgaagtcg aggcatttct gtcctggctg gcgaacgagc gcaaggtttc
     2881 ggtctccacg accggtatcc cgccgccgcc gccggatacc ggttaatagt acctgtgaag
     2941 tgaaaaatgg cgcacattgt gcgacatttt tttctcgagc aggtagtcgg ccaataaccc
     3001 agggattagt ctgctcgagg cgatcgagat cgtagaaccg accgggttca acccagagtt
     3061 gcttaccgct tgcttcgatg attgcgtaag ccatgcattt tcagccgtag aggtggctac
     3121 acctgccgag ggcagagaga gcacttgaag aaacctgttc cgagccgcct tttaaacaac
     3181 agccctctat catctcagaa acttttgaga actgtcaaac tgtggacaaa gcgatcgcgg
     3241 cctcatgagc tgcagtgcta ggctaaatta aatttttcct ttgtccagag attaatctgt
     3301 ctgcagactc agtcctgaca ggagcgactg cgtgctctcg caaattgcaa tctgcatttg
     3361 ggtggaatcg acggcaattt tgcaggattg ccagcgggcg ctgtcggccg cacgagaaac
     3421 ctgaaaaggt aaaggaggtc ttaagctcgg ctcaatttct ctctttatcc tgttagatgg
     3481 tttgattgct gttgctaccc cgttgatctg cgtatgagcc ctcgtaaaac ctacattctc
     3541 aagctctacg tcgccggcga acttcaagat tccgacgact tctaactgtc gttaactgct
     3601 ttgttggtac tacctgactt caccctcttt taagatgact tccgctgaga tgactagccc
     3661 taataataat tctgagcacc aagccatcgc taagatgcgc acgatgattg aaggctttga
     3721 tgatattagt catggcggtc ttcctatcgg gcgatcgacc ctcgttagtg gtacttcagg
     3781 aaccggcaag accctttttt ctattcaatt tctctataac ggtattatcg agtttgatga
     3841 gcctggggtt ttcgttactt tcgaagaaac cccgcaagat atcattaaaa acgcccgtag
     3901 ttttggctgg gatttagcca agctggtcga tgagggcaaa ctatttattc ttgatgcttc
     3961 acccgatcca gaaggtcaag aggttgttgg cggcttcgat ctctctgctc tgattgagcg
     4021 gattaattat gcaattcaaa agtatcgagc gcggcgggtt tcaattgact cggtcacgtc
     4081 cgttttccag caatatgatg cctcttctgt ggttcgccgc gaactctttc ggttggtagc
     4141 tcgcctaaaa caaattgggg caactacggt catgaccacc gagcgtatcg aggaatatgg
     4201 cccgatcgct cgttacggtg ttgaggaatt tgtctccgat aacgtcgtga ttctccgcaa
     4261 cgttttggaa ggggagcgcc gtcgccgcac cctcgaaatc ctcaagctac gtggcaccag
     4321 ccacatgaaa ggggaatatc cgttcacgat tacggatcat ggcatcaata tcttcccgct
     4381 cggggcaatg cgccttacgc agcgatcgtc gaacgtgcgt gtttcatctg gtgtcgtccg
     4441 actcgatgaa atgtgtggtg ggggcttctt taaggactca atcattctgg caactggcgc
     4501 tacaggcact ggtaaaactc tgttagttag ccgtttcgtt gagaatgctt gtgctaacaa
     4561 agagcgggcg attctgttcg cttatgaaga gtcacgagct cagctgctcc gcaatgccta
     4621 ttcatgggga atggactttg aggagatgga gcgccaaaac ctcctcaaaa ttgtttgcgc
     4681 ctatcctgaa tctgcaggtc ttgaagacca tttgcagatt attaagtcgg agatcaatga
     4741 ctttaagcca gctcgtattg caatcgactc cctctctgct ttggcgcggg gcgttagcaa
     4801 caatgccttc cgccaatttg taattggtgt cactggctac gcgaaacaag aagaaatcac
     4861 gggactattc acaaatacca gtgatcaatt tatgggagcg cattcgatta ctgactccca
     4921 tatctcaaca attacggata cgattatctt gctccaatac gtcgagattc gtggcgaaat
     4981 gtcccgcgcc attaacgtct tcaagatgcg cggatcttgg catgacaaag caatccgcga
     5041 attcatgatc agcgacaaag ggccggacat caaggattct ttccggaact ttgagcggat
     5101 tatttcaggt tcgccaacac ggattaccgt cgatgagaaa agcgaactct cgcgaattgt
     5161 gcgcggcgtt caagaaaaag ggccggagag cggctcgagt ggatccgcaa gcagtggggc
     5221 cggcggagct ggagcagcta gcggaagtgc aggtgaactt gctgttgaag ctaagcttga
     5281 agttgctatg gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga
     5341 gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc
     5401 cacctacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg
     5461 gcccaccctc gtgaccaccc tgacctgggg cgtgcagtgc ttcagccgct accccgacca
     5521 catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac
     5581 catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga
     5641 caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct
     5701 ggggcacaag ctggagtaca actacatcag ccacaacgtc tatatcaccg ccgacaagca
     5761 gaagaacggc atcaaggcca acttcaagat ccgccacaac atcgaggacg gcagcgtgca
     5821 gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga
     5881 caaccactac ctgagcaccc agtccgccct gagcaaagac cccaacgaga agcgcgatca
     5941 catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta
     6001 caagtaaaaa tgaaggagct ccccggcagg agaaggccat cctgacggat ggcctttttg
     6061 aagctttcta gaatcggggg cccccggggg gattctagag ttgctaccaa gtagtgaccc
     6121 gcttcgtgat gcaaaatccg ctgacgatat tcgggcgatc gctgctgaat gccatcgagc
     6181 agtaacgtgg caccccgccc ctgccaagtc accgcatcca gactgaacag caccaagagg
     6241 ctaaaaccca atcccgccgg tagcagcgga gaactaccca gcattggtcc caccaaagct
     6301 aatgccgtcg tggtaaaaat cgcgatcgcc gtcagactca agcccagttc gctcatgctt
     6361 cctcatctag gtcacagtct tcggcgatcg catcgatctg atgctgcagc aagcgttttc
     6421 cataccggcg atcgcgccgt cgccctttcg ctgccgtggc ccgcttacga gctcgtttat
     6481 cgaccacgat cgcatccaaa tccgcgatcg cttcccagtc cggcaattca gtctggggcg
     6541 tccgtttcat taatcctgat agcgtcagac cccgtagaaa agatcaaag
//
