LOCUS       sequence-95265-e        4296 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Constitutively expresses eGFP with proB promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4296)
  AUTHORS   Bienick MS, Young KW, Klesmith JR, Detwiler EE, Tomek KJ, Whitehead 
            TA
  TITLE     The Interrelationship between Promoter Strength, Gene Expression, 
            and Growth Rate.
  JOURNAL   PLoS One. 2014 Oct 6;9(10):e109105. doi: 
            10.1371/journal.pone.0109105. eCollection 2014.
  PUBMED    25286161
REFERENCE   2  (bases 1 to 4296)
  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: "PLoS One.";
            date: "2014-10-6"; volume: "9(10):e109105. doi"; pages: " 
            10.1371/journal.pone.0109105. eCollection 2014"
COMMENT     Sequence Label: pJK_proB_eGFP
FEATURES             Location/Qualifiers
     source          1..4296
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      12..467
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(99..118)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     309..330
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(join(560..1351,1352..1420))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     1183..1202
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1542..2130
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2031..2050
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2284..2301
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2316..2458
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2384..2403)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2544..2566
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2560..2751)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     RBS             3390..3412
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             3420..4133
                     /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"
     CDS             3420..4133
                     /codon_start=1
                     /product="Aequorea victoria green fluorescent protein"
                     /label=GFP
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(3450..3478)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     4073..4093
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     CDS             4140..4157
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4210..4228)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      4224..4271
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
ORIGIN
        1 tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
       61 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
      121 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
      181 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
      241 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
      301 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
      361 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
      421 acaaaaattt aacgcgaatt ttaacaaaat attaacgttt acaatttcag gtggcacttt
      481 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
      541 tccgctcatg aattaattct taccaatgct taatcagtga ggcacctatc tcagcgatct
      601 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
      661 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
      721 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
      781 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
      841 cagttaatag tttgcgcaac gttgttgcca ttgctgcagg catcgtggtg tcacgctcgt
      901 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
      961 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     1021 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     1081 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     1141 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     1201 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     1261 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     1321 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     1381 aaaagggaat aagggcgaca cggaaatgtt gaatactcat aacacccctt gtattactgt
     1441 ttatgtaagc agacagtttt attgttcatg accaaaatcc cttaacgtga gttttcgttc
     1501 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     1561 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     1621 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     1681 aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     1741 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     1801 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     1861 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     1921 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     1981 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     2041 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     2101 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     2161 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     2221 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
     2281 cgcagcgagt cagtgagcga ggaagcggaa gagcgcctga tgcggtattt tctccttacg
     2341 catctgtgcg gtatttcaca ccgcatatat ggtgcactct cagtacaatc tgctctgatg
     2401 ccgcatagtt aagccagtat acactccgct atcgctacgt gactgggtca tggctgcgcc
     2461 ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc
     2521 ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc
     2581 accgaaacgc gcgaggcagc tgcggtaaag ctcatcagcg tggtcgtgaa gcgattcaca
     2641 gatgtctgcc tgttcatccg cgtccagctc gttgagtttc tccagaagcg ttaatgtctg
     2701 gcttctgata aagcgggcca tgttaagggc ggttttttcc tgtttggtca ctgatgcctc
     2761 cgtgtaaggg ggatttctgt tcatgggggt aatgataccg atgaaacgag agaggatgct
     2821 cacgatacgg gttactgatg atgaacatgc ccggttactg gaacgttgtg agggtaaaca
     2881 actggcggta tggatgcggc gggaccagag aaaaatcact cagggtcaat gccagcgctt
     2941 cgttaataca gatgtaggtg ttccacaggg tagccagcag catcctgcga tgcagatccg
     3001 gaacataatg gtgcagggcg ctgacttccg cgtttccaga ctttacgaaa cacggaaacc
     3061 gaagaccatt catgttgttg ctcaggtcgc agacgttttg cagcagcagt cgcttcacgt
     3121 tcgctcgcgt atcggtgatt cattctgcta accagtaagg caaccccgcc agcctagccg
     3181 ggtcctcaac gacaggagca cgatcatgcg cacccgtggg gccgccatgc cggccacagc
     3241 taacaccacg tcgtccctat ctgctgccct aggtctatga gtggttgctg gataacttta
     3301 cgggcatgca taaggctcgt aatatatatt cagggagacc acaacggttt ccctctacaa
     3361 ataattttgt ttaactttct agaaataatt ttgtttaact ttaagaagga gatatacata
     3421 tgcgtaaagg agaagaactt ttcactggag ttgtcccaat tcttgttgaa ttagatggtg
     3481 atgttaatgg gcacaaattt tctgtcagtg gagagggtga aggtgatgca acatacggaa
     3541 aacttaccct taaatttatt tgcactactg gaaaactacc tgttccatgg ccaacacttg
     3601 tcactacttt cggttatggt gttcaatgct ttgcgagata cccagatcat atgaaacagc
     3661 atgacttttt caagagtgcc atgcccgaag gttatgtaca ggaaagaact atatttttca
     3721 aagatgacgg gaactacaag acacgtgctg aagtcaagtt tgaaggtgat acccttgtta
     3781 atagaatcga gttaaaaggt attgatttta aagaagatgg aaacattctt ggacacaaat
     3841 tggaatacaa ctataactca cacaatgtat acatcatggc agacaaacaa aagaatggaa
     3901 tcaaagttaa cttcaaaatt agacacaaca ttgaagatgg aagcgttcaa ctagcagacc
     3961 attatcaaca aaatactcca attggcgatg gccctgtcct tttaccagac aaccattacc
     4021 tgtccacaca atctgccctt tcgaaagatc ccaacgaaaa gagagaccac atggtccttc
     4081 ttgagtttgt aacagctgct gggattacac atggcatgga tgaactatac aaactcgagc
     4141 accaccacca ccaccactga gatccggctg ctaacaaagc ccgaaaggaa gctgagttgg
     4201 ctgctgccac cgctgagcaa taactagcat aaccccttgg ggcctctaaa cgggtcttga
     4261 ggggtttttt gctgaaagga ggaactatat ccggat
//
