LOCUS       sequence-256213-        4480 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  express GFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4480)
  TITLE     Emory Custom Cloning Core Plasmids - Oskar Laur
REFERENCE   2  (bases 1 to 4480)
  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"
COMMENT     Sequence Label: pTRE2 EGFP
FEATURES             Location/Qualifiers
     source          1..4480
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    join(20..38,39..61,62..80,81..103,104..122,123..145,
                     146..164,165..187,188..206,207..229,230..248,249..271,
                     272..290)
                     /label=tetracycline response element
                     /bound_moiety="tetracycline repressor TetR"
                     /note="contains seven copies of the tetracycline operator 
                     tetO"
     promoter        323..361
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     358..382
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      489..498
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(495..497,498..500,501..1214)
                     /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(495..497,498..500,501..1211)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(540..561)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(801..820)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1148..1169
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     intron          1269..1841
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     primer_bind     complement(1973..1992)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2038..2093
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2092..2111)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(2491..2508)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2662..3250)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2742..2761)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3421..4212,4213..4281))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     4044..4063
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4282..4386)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4454..4472
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 cttcactcga gtttaccact ccctatcagt gatagagaaa agtgaaagtc gagtttacca
       61 ctccctatca gtgatagaga aaagtgaaag tcgagtttac cactccctat cagtgataga
      121 gaaaagtgaa agtcgagttt accactccct atcagtgata gagaaaagtg aaagtcgagt
      181 ttaccactcc ctatcagtga tagagaaaag tgaaagtcga gtttaccact ccctatcagt
      241 gatagagaaa agtgaaagtc gagtttacca ctccctatca gtgatagaga aaagtgaaag
      301 tcgagctcgg tacccgggtc gaggtaggcg tgtacggtgg gaggcctata taagcagagc
      361 tcgtttagtg aaccgtcaga tcgcctggag acgccatcca cgctgttttg acctccatag
      421 aagacaccgg gaccgatcca gcctccgcgg ccccgaattc gagctcggta cccggggatc
      481 caccggtcgc caccatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc
      541 tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg
      601 gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg
      661 tgccctggcc caccctcgtg accaccctga cctacggcgt gcagtgcttc agccgctacc
      721 ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg
      781 agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg
      841 agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca
      901 acatcctggg gcacaagctg gagtacaact acaacagcca caacgtctat atcatggccg
      961 acaagcagaa gaacggcatc aaggtgaact tcaagatccg ccacaacatc gaggacggca
     1021 gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc
     1081 tgcccgacaa ccactacctg agcacccagt ccgccctgag caaagacccc aacgagaagc
     1141 gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg
     1201 agctgtacaa gtaaagcggc cgcatcgata agcttgtcga cgatatctct agagctgaga
     1261 acttcagggt gagtttgggg acccttgatt gttctttctt tttcgctatt gtaaaattca
     1321 tgttatatgg agggggcaaa gttttcaggg tgttgtttag aatgggaaga tgtcccttgt
     1381 atcaccatgg accctcatga taattttgtt tctttcactt tctactctgt tgacaaccat
     1441 tgtctcctct tattttcttt tcattttctg taactttttc gttaaacttt agcttgcatt
     1501 tgtaacgaat ttttaaattc acttttgttt atttgtcaga ttgtaagtac tttctctaat
     1561 cacttttttt tcaaggcaat cagggtatat tatattgtac ttcagcacag ttttagagaa
     1621 caattgttat aattaaatga taaggtagaa tatttctgca tataaattct ggctggcgtg
     1681 gaaatattct tattggtaga aacaactaca ccctggtcat catcctgcct ttctctttat
     1741 ggttacaatg atatacactg tttgagatga ggataaaata ctctgagtcc aaaccgggcc
     1801 cctctgctaa ccatgttcat gccttcttct ctttcctaca gctcctgggc aacgtgctgg
     1861 ttgttgtgct gtctcatcat tttggcaaag aattcactcc tcaggtgcag gctgcctatc
     1921 agaaggtggt ggctggtgtg gccaatgccc tggctcacaa ataccactga gatctttttc
     1981 cctctgccaa aaattatggg gacatcatga agccccttga gcatctgact tctggctaat
     2041 aaaggaaatt tattttcatt gcaatagtgt gttggaattt tttgtgtctc tcactcggaa
     2101 ggacatatgg gagggcaaat catttaaaac atcagaatga gtatttggtt tagagtttgg
     2161 caacatatgc ccatatgctg gctgccatga acaaaggttg gctataaaga ggtcatcagt
     2221 atatgaaaca gccccctgct gtccattcct tattccatag aaaagccttg acttgaggtt
     2281 agattttttt tatattttgt tttgtgttat ttttttcttt aacatcccta aaattttcct
     2341 tacatgtttt actagccaga tttttcctcc tctcctgact actcccagtc atagctgtcc
     2401 ctcttctctt atggagatcc ctcgactgca ttaatgaatc ggccaacgcg cggggagagg
     2461 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     2521 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     2581 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     2641 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     2701 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     2761 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     2821 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     2881 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     2941 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     3001 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     3061 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     3121 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     3181 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     3241 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     3301 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     3361 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     3421 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     3481 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     3541 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     3601 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     3661 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     3721 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     3781 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     3841 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     3901 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     3961 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     4021 ctcttgcccg gcgtcaacac gggataatac cgcgccacat agcagaactt taaaagtgct
     4081 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     4141 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     4201 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     4261 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     4321 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     4381 tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac
     4441 attaacctat aaaaataggc gtatcacgag gccctttcgt
//
