LOCUS       sequence-373903-        3210 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses EGFP from PlLaco-1 promoter in E.coli Dh5alpha cell. For 
            experiment use Dh5alphaz1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3210)
  AUTHORS   Mukhopadhyay S, Bagh S
  TITLE     A microgravity responsive synthetic genetic device in Escherichia 
            coli.
  JOURNAL   Biosens Bioelectron. 2020 Nov 1;167:112462. doi: 
            10.1016/j.bios.2020.112462. Epub 2020 Aug 1.
  PUBMED    32781386
REFERENCE   2  (bases 1 to 3210)
  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: "Biosens 
            Bioelectron."; date: "2020-11-1"; volume: "167:112462. doi"; pages: 
            " 10.1016/j.bios.2020.112462. Epub 2020 Aug 1"
COMMENT     Sequence Label: L_EGFP_AC
FEATURES             Location/Qualifiers
     source          1..3210
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    22..38
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     protein_bind    45..61
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     RBS             103..114
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             join(121..123,124..126,127..840)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(121..123,124..126,127..837)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(166..187)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(427..446)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     774..795
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     terminator      864..950
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(985..1002)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1119..1664)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     complement(1199..1218)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(1778..1872)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(1893..2552)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     2467..2486
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(2553..2655)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     2721..2739
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 caaaagcact ctgctcgaga attgtgagcg gataacaatt gacattgtga gcggataaca
       61 agatactgag cacatcagca ggacgcactg accgaattca ttaaagagga gaaaggtacc
      121 atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac
      181 ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac
      241 ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc
      301 ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc gctaccccga ccacatgaag
      361 cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc
      421 ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg
      481 gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac
      541 aagctggagt acaactacaa cagccacaac gtctatatca tggccgacaa gcagaagaac
      601 ggcatcaagg tgaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc
      661 gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac
      721 tacctgagca cccagtccgc cctgagcaaa gaccccaacg agaagcgcga tcacatggtc
      781 ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagtaa
      841 agcggccgcg actctagagg catcaaataa aacgaaaggc tcagtcgaaa gactgggcct
      901 ttcgttttat ctgttgtttg tcggtgaacg ctctcctgag taggacaaat ccgccgccct
      961 agacctaggg atatattccg cttcctcgct cactgactcg ctacgctcgg tcgttcgact
     1021 gcggcgagcg gaaatggctt acgaacgggg cggagatttc ctggaagatg ccaggaagat
     1081 acttaacagg gaagtgagag ggccgcggca aagccgtttt tccataggct ccgcccccct
     1141 gacaagcatc acgaaatctg acgctcaaat cagtggtggc gaaacccgac aggactataa
     1201 agataccagg cgtttccccc tggcggctcc ctcgtgcgct ctcctgttcc tgcctttcgg
     1261 tttaccggtg tcattccgct gttatggccg cgtttgtctc attccacgcc tgacactcag
     1321 ttccgggtag gcagttcgct ccaagctgga ctgtatgcac gaaccccccg ttcagtccga
     1381 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggaaagac atgcaaaagc
     1441 accactggca gcagccactg gtaattgatt tagaggagtt agtcttgaag tcatgcgccg
     1501 gttaaggcta aactgaaagg acaagttttg gtgactgcgc tcctccaagc cagttacctc
     1561 ggttcaaaga gttggtagct cagagaacct tcgaaaaacc gccctgcaag gcggtttttt
     1621 cgttttcaga gcaagagatt acgcgcagac caaaacgatc tcaagaagat catcttatta
     1681 atcagataaa atatttctag atttcagtgc aatttatctc ttcaaatgta gcacctgaag
     1741 tcagccccat acgatataag ttgttactag tgcttggatt ctcaccaata aaaaacgccc
     1801 ggcggcaacc gagcgttctg aacaaatcca gatggagttc tgaggtcatt actggatcta
     1861 tcaacaggag tccaagcgag ctcgatatca aattacgccc cgccctgcca ctcatcgcag
     1921 tactgttgta attcattaag cattctgccg acatggaagc catcacaaac ggcatgatga
     1981 acctgaatcg ccagcggcat cagcaccttg tcgccttgcg tataatattt gcccatggtg
     2041 aaaacggggg cgaagaagtt gtccatattg gccacgttta aatcaaaact ggtgaaactc
     2101 acccagggat tggctgagac gaaaaacata ttctcaataa accctttagg gaaataggcc
     2161 aggttttcac cgtaacacgc cacatcttgc gaatatatgt gtagaaactg ccggaaatcg
     2221 tcgtggtatt cactccagag cgatgaaaac gtttcagttt gctcatggaa aacggtgtaa
     2281 caagggtgaa cactatccca tatcaccagc tcaccgtctt tcattgccat acggaattcc
     2341 ggatgagcat tcatcaggcg ggcaagaatg tgaataaagg ccggataaaa cttgtgctta
     2401 tttttcttta cggtctttaa aaaggccgta atatccagct gaacggtctg gttataggta
     2461 cattgagcaa ctgactgaaa tgcctcaaaa tgttctttac gatgccattg ggatatatca
     2521 acggtggtat atccagtgat ttttttctcc attttagctt ccttagctcc tgaaaatctc
     2581 gataactcaa aaaatacgcc cggtagtgat cttatttcat tatggtgaaa gttggaacct
     2641 cttacgtgcc gatcaacgtc tcattttcgc cagatatcga cgtctaagaa accattatta
     2701 tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtctt cacttgtttt
     2761 ggagcacgga aagacgatga cggaaaaaga gatcgtggat tacgtcgcca gtcaagtaac
     2821 aaccgccaaa aagttgcgcg gaggagttgt gtttgtggac gaagtaccga aaggtcttac
     2881 cggaaaactc gacgcaagaa aaatcagaga gatcctcata aaggccaaga aggatcctgg
     2941 cagaagctat gaaattcttt atgccggctg cagcagattc tcgcatgcca gagatcctat
     3001 ttttggcaat caaatcattc cggatactgc gattttaagt gttgttccat tccatcacgg
     3061 ttttggaatg tttactacac tcggatattt gatatgtgga tttcgagtcg tcttaatgta
     3121 tagatttgaa gaagagctgt ttttacgatc ccttcaggat tacaaaattc aaagtgcgtt
     3181 gctagtacca accctatttt cattcttcgc
//
