LOCUS       sequence-173839-        9369 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  BbbJ23119::EYFP cloned between XbaI and XhoI of EB2065.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9369)
  AUTHORS   Zheng XY, O'Shea EK
  TITLE     Cyanobacteria Maintain Constant Protein Concentration despite Genome
            Copy-Number Variation.
  JOURNAL   Cell Rep. 2017 Apr 18;19(3):497-504. doi: 
            10.1016/j.celrep.2017.03.067.
  PUBMED    28423314
REFERENCE   2  (bases 1 to 9369)
  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.celrep.2017.03"; journalName: "Cell Rep"; date: 
            "2017-04-18- 18"; volume: "19"; issue: "3"; pages: "497-504"
COMMENT     Sequence Label: EB2315
FEATURES             Location/Qualifiers
     source          1..9369
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1111..1145
                     /label=J23119(SpeI) promoter
                     /note="bacterial promoter (Registry of Standard Biological 
                     Parts BBa_J23119) modified to end with an SpeI site"
     CDS             join(1160..1162,1163..1165,1166..1879)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVRFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1205..1226)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1466..1485)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1813..1834
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     terminator      complement(1935..1962)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(2054..2140)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     promoter        complement(2718..2822)
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3025..3127
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             3128..3787
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(3194..3213)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     primer_bind     complement(5071..5090)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             6548..6739
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(6733..6755)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    6841..6981
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     6896..6915
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(6996..7013)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7167..7755)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7247..7266)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7926..8717,8718..8786))
                     /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(8787..8891)
                     /gene="bla"
                     /label=AmpR promoter
     promoter        join(9004..9009,9010..9026,9027..9032)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
ORIGIN
        1 gatcccaggc tcagtgtggt tcgcatcttt tgataatcac cactggacgc aacacagtag
       61 ggcaattcaa gagcatccaa agcctcttca accccaggaa cggcttgtag atgcgtttct
      121 aacgcgatac gggtacggcg ttgatagtgc tgaacaaagt cagggggtgg aggattgcct
      181 aaccgtcgct caattagttt gagacagtca gccatggaat gacccacaaa ctgctcaaac
      241 atgtcatcca aagtcaccaa cagacccagt tcattgagca tgtctgcaaa gacgcgatta
      301 gtgatgcgtt cgctatcaac aagcacacca tcacagtcga aaatcaccag ctgaaacggt
      361 gaagtttgca ttgtttttaa gcacgagcca tcaacagtaa gagcgatcgc gctgggacga
      421 tgtaatcgcg ccgtaggcag ggttttgcct catccggcag atgacaagct tcaagattcg
      481 gcagtgaagt atcgagcaag cgataccaaa cgcggccgcg aggaggcctc ggcaactgga
      541 agcgcaggtc ttcccagtaa gcattaaagg ctaggtaaag ccattcctgc tggcgaggat
      601 ggcagagact gacggccaga ctgtgggacc acagcgccca atcgggttgt ttgagtttga
      661 cgccatgcca gatggcatag ggacgacgcg gatggggttc gttctgcagc agttcgttct
      721 gttggaacat caccagcgac tgggaaagtt caatcaggcg gcgactgaac accaagaaat
      781 cggcatggcg atcgcacagc gaccaatcaa accagctgat ctcattgtct tggcagtagg
      841 cgttattgtt accctgctga ctgcgtttga cctcatcgcc catcgtcagc atcggtgtgc
      901 cctgggcgag gaataacgtg gcgagcaaat tgcgctgctg ccgttcccgt aagctcagaa
      961 tcgtggggtc atcggtctcg ccttcaatgc cgtagttcca gctgtagttg tcattggtcc
     1021 cgtcccgatt gttctctcca ttggcaaagt tgtgcttctg gctatagctg actcgtcacg
     1081 ggtaaccgat atcgtcgaca ggcctctaga ttgacagcta gctcagtcct aggtataata
     1141 ctagtaagga ggcaacaaga tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc
     1201 catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg
     1261 cgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct
     1321 gcccgtgccc tggcccaccc tcgtgaccac cttcggctac ggcctgcagt gcttcgcccg
     1381 ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt
     1441 ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgag
     1501 gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga
     1561 cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg tctatatcat
     1621 ggccgacaag cagaagaacg gcatcaaggt gaacttcaag atccgccaca acatcgagga
     1681 cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt
     1741 gctgctgccc gacaaccact acctgagcta ccagtccgcc ctgagcaaag accccaacga
     1801 gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat
     1861 ggacgagctg tacaagtaac tcgagctagc aagcttggcc ggatccggcc ggatccggag
     1921 tttgtagaaa cgcaaaaagg ccatccgtca ggatggcctt ctgcttaatt tgatgcctgg
     1981 cagtttatgg cgggcgtcct gcccgccacc ctccgggccg ttgcttcgca acgttcaaat
     2041 ccgctcccgg cggatttgtc ctactcagga gagcgttcac cgacaaacaa cagataaaac
     2101 gaaaggccca gtctttcgac tgagcctttc gttttatttg atgcctggca gttccctact
     2161 ctcgcatggg gagaccccac actaccatcg gcgctacggc gtttcacttc tgagttcggc
     2221 atggggtcag gtgggaccac cgcgctactg ccgccaggca aattctgttt tatcagccgt
     2281 taccccacct actagctaat cccatctggg cacatccgat ggcaagaggc ccgaaggtcc
     2341 ccctctttgg tcttgcgacg ttatgcggta ttagctaccg tttccagtag ttatccccct
     2401 ccatcaggca gtttcccaga cattactcac ccgtccgcca ctcgtcagca aagaagcaag
     2461 cttagatccg tcacacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     2521 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     2581 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     2641 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     2701 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     2761 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     2821 cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta
     2881 acctataaaa ataggcgtat cacgaggccc tttcgtcttc gaataaatac ctgtgacgga
     2941 agatcacttc gcagaataaa taaatcctgg tgtccctgtt gataccggga agccctgggc
     3001 caacttttgg cgaaaatgag acgttgatcg gcacgtaaga ggttccaact ttcaccataa
     3061 tgaaataaga tcactaccgg gcgtattttt tgagttatcg agattttcag gagctaagga
     3121 agctaaaatg gagaaaaaaa tcactggata taccaccgtt gatatatccc aatggcatcg
     3181 taaagaacat tttgaggcat ttcagtcagt tgctcaatgt acctataacc agaccgttca
     3241 gctggatatt acggcctttt taaagaccgt aaagaaaaat aagcacaagt tttatccggc
     3301 ctttattcac attcttgccc gcctgatgaa tgctcatccg gaattccgta tggcaatgaa
     3361 agacggtgag ctggtgatat gggatagtgt tcacccttgt tacaccgttt tccatgagca
     3421 aactgaaacg ttttcatcgc tctggagtga ataccacgac gatttccggc agtttctaca
     3481 catatattcg caagatgtgg cgtgttacgg tgaaaacctg gcctatttcc ctaaagggtt
     3541 tattgagaat atgtttttcg tctcagccaa tccctgggtg agtttcacca gttttgattt
     3601 aaacgtggcc aatatggaca acttcttcgc ccccgttttc accatgggca aatattatac
     3661 gcaaggcgac aaggtgctga tgccgctggc gattcaggtt catcatgccg tttgtgatgg
     3721 cttccatgtc ggcagaatgc ttaatgaatt acaacagtac tgcgatgagt ggcagggcgg
     3781 ggcgtaattt ttttaaggca gttattggtg cccttaaacg cctggttgct acgcctgaat
     3841 aagtgataat aagcggatga atggcagaaa ttcgaaagca aattcgaccc ggtcgtcggt
     3901 tcagggcagg gtcgttaaat agccgcttat gtctattgct ggtttaccgg tttattgact
     3961 accggaagca gtgtgaccgt gtgcttctca aatgcctgag gccagtttgc tcaggctctc
     4021 cccgtggagg taataattga cgatatgatc gacggatctg gtaaccccag cgcggagatc
     4081 tcgcagcgta aagccgtcat ggcaggtgat gaagttaatg gtgcgtccgg cataccattg
     4141 gtctgtgctg tagacatcgg ggctacccag caggcgttga ctgagggcgt aagtacagcc
     4201 ctgatctcca cgccaaaaac gccgaatatc gtcccggaag ggaccgttcc aagtcccaaa
     4261 gcgatcgcca ataaaggtac caacctgata taagccggct gcgtcccaag cttcagcaat
     4321 gagcttcgta ccggccaaaa ccggatcgga atcaatcgcc caaagcaagg gcggatccga
     4381 tagggggttg ccattggcat cacgactcag caccgacgca aggtcaaagc ggaagccatc
     4441 gacgtgcatt tccgagaccc aataacgcag gcaatcgaga atcagcgatc gcacgatcga
     4501 attgttagct ttgacggtgt tgccgcagcc ggtgtagttg cgatagcccg cctgttcatc
     4561 cagcagatag taggttgaat tcgctagacc tttgaaagac agcgttggac cgtcttcatt
     4621 cccttcagca gtgtggttga acaccacgtc gagaatcacc tcaatccctg cttggtggag
     4681 cgccttgacg aggtcgcgga actcatcaac tggaccaagt ggatcatggc gagagctgta
     4741 ggctgcgtgg ggcgcaaaaa aagccatggt gctgtagccc cagtagttct cgcgtcctaa
     4801 gggggcatct tggcgatcga actggtgcac cggcagcaac tcaacggccg tgacgccgag
     4861 ggattgcagg tagggaattt tttcgattag accagcgtag gtgccacgtt tttcaggggc
     4921 gacgccggaa ttgggatgct tggtgaagcc gccaacatgc agctcataga ttactgtgcg
     4981 agcgtagggt gtgcgtggat ggcgatcgga tcctctacgc cggacgcatc gtggccggca
     5041 tcaccggcgc cacaggtgcg gttgctggcg cctatatcgc cgacatcacc gatggggaag
     5101 atcgggctcg ccacttcggg ctcatgagcg cttgtttcgg cgtgggtatg gtggcaggcc
     5161 ccgtggccgg gggactgttg ggcgccatct ccttgcatgc accattcctt gcggcggcgg
     5221 tgctcaacgg cctcaaccta ctactgggct gcttcctaat gcaggagtcg cataagggag
     5281 agcgtcgacc gatgcccttg agagccttca acccagtcag ctccttccgg tgggcgcggg
     5341 gcatgactat cgtcgccgca cttatgactg tcttctttat catgcaactc gtaggacagg
     5401 tgccggcagc gctctgggtc attttcggcg aggaccgctt tcgctggagc gcgacgatga
     5461 tcggcctgtc gcttgcggta ttcggaatct tgcacgccct cgctcaagcc ttcgtcactg
     5521 gtcccgccac caaacgtttc ggcgagaagc aggccattat cgccggcatg gcggccgacg
     5581 cgctgggcta cgtcttgctg gcgttcgcga cgcgaggctg gatggccttc cccattatga
     5641 ttcttctcgc ttccggcggc atcgggatgc ccgcgttgca ggccatgctg tccaggcagg
     5701 tagatgacga ccatcaggga cagcttcaag gatcgctcgc ggctcttacc agcctaactt
     5761 cgatcattgg accgctgatc gtcacggcga tttatgccgc ctcggcgagc acatggaacg
     5821 ggttggcatg gattgtaggc gccgccctat accttgtctg cctccccgcg ttgcgtcgcg
     5881 gtgcatggag ccgggccacc tcgacctgaa tggaagccgg cggcacctcg ctaacggatt
     5941 caccactcca agaattggag ccaatcaatt cttgcggaga actgtgaatg cgcaaaccaa
     6001 cccttggcag aacatatcca tcgcgtccgc catctccagc agccgcacgc ggcgcatctc
     6061 gggcagcgtt gggtcctggc cacgggtgcg catgatcgtg ctcctgtcgt tgaggacccg
     6121 gctaggctgg cggggttgcc ttactggtta gcagaatgaa tcaccgatac gcgagcgaac
     6181 gtgaagcgac tgctgctgca aaacgtctgc gacctgagca acaacatgaa tggtcttcgg
     6241 tttccgtgtt tcgtaaagtc tggaaacgcg gaagtcagcg ccctgcacca ttatgttccg
     6301 gatctgcatc gcaggatgct gctggctacc ctgtggaaca cctacatctg tattaacgaa
     6361 gcgctggcat tgaccctgag tgatttttct ctggtcccgc cgcatccata ccgccagttg
     6421 tttaccctca caacgttcca gtaaccgggc atgttcatca tcagtaaccc gtatcgtgag
     6481 catcctctct cgtttcatcg gtatcattac ccccatgaac agaaatcccc cttacacgga
     6541 ggcatcagtg accaaacagg aaaaaaccgc ccttaacatg gcccgcttta tcagaagcca
     6601 gacattaacg cttctggaga aactcaacga gctggacgcg gatgaacagg cagacatctg
     6661 tgaatcgctt cacgaccacg ctgatgagct ttaccgcagc tgcctcgcgc gtttcggtga
     6721 tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc
     6781 ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg
     6841 cgcagccatg acccagtcac gtagcgatag cggagtgtat actggcttaa ctatgcggca
     6901 tcagagcaga ttgtactgag agtgcaccat atgcggtgtg aaataccgca cagatgcgta
     6961 aggagaaaat accgcatcag gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     7021 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     7081 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     7141 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     7201 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     7261 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     7321 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     7381 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     7441 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     7501 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     7561 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     7621 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     7681 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     7741 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     7801 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     7861 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     7921 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     7981 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     8041 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     8101 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     8161 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     8221 cgcaacgttg ttgccattgc tgcaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     8281 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     8341 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     8401 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     8461 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     8521 agttgctctt gcccggcgtc aacacgggat aataccgcgc cacatagcag aactttaaaa
     8581 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     8641 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     8701 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     8761 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     8821 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     8881 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac cattattatc
     8941 atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtcttca agaattctca
     9001 tgtttgacag cttatcatcg ataagcttta atgcggtagt ttatcacagt taaattgcta
     9061 acgcagtcag gcaccgtgta tgaaatctaa caatgcgctc atcgtcatcc tcggcaccgt
     9121 caccctggat gctgtaggca taggcttggt tatgccggta ctgccgggcc tcttgcggga
     9181 tatcgtccat tccgacagca tcgccagtca ctatggcgtg ctgctagcgc tatatgcgtt
     9241 gatgcaattt ctatgcgcac ccgttctcgg agcactgtcc gaccgctttg gccgccgccc
     9301 agtcctgctc gcttcgctac ttggagccac tatcgactac gcgatcatgg cgaccacacc
     9361 cgtcctgtg
//
