LOCUS       sequence-378936-        6013 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expression of EGFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6013)
  TITLE     Oka Lab Plasmids 2022
REFERENCE   2  (bases 1 to 6013)
  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: scAV0.1-TBG-EGFP
FEATURES             Location/Qualifiers
     source          1..6013
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        327..736
                     /label=TBG promoter
                     /note="promoter from the human thyroxine-binding globulin 
                     gene (Yan et al., 2012)"
     CDS             join(776..778,779..781,782..1495)
                     /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(776..778,779..781,782..1492)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(821..842)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1082..1101)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1429..1450
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1599..1619)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     primer_bind     complement(1807..1824)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1813..2020
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2033..2173
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2033..2162
                     /label=AAV2 ITR
     primer_bind     2980..3002
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3040..3058)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3126..3230
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3231..3299,3300..4091)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(3449..3468)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4262..4850
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     rep_origin      5457..5970
                     /direction=RIGHT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5544..5563)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5754..5775
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 ctgcgcgctc gctcgctcac tgaggccgcc cgggcaaagc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggaatt cacgcgtgga
      121 tctgaattca attcacgcgt ggtacgtgga gtcgtgacgt gaattacgtc atagggttag
      181 ggaggtcctg tattagaggt cacgtgagtg ttttgcgaca ttttgcgaca ccatgtggtc
      241 acgctgggta tttaagcccg agtgagcacg cagggtctcc attgctaggg gctggaagct
      301 acctttgaca tcatttcctc tgcgaatgca tgtataattt ctacagaacc tattagaaag
      361 gatcacccag cctctgcttt tgtacaactt tcccttaaaa aactgccaat tccactgctg
      421 tttggcccaa tagtgagaac tttttcctgc tgcctcttgg tgcttttgcc tatggcccct
      481 attctgcctg ctgaagacac tcttgccagc atggacttaa acccctccag ctctgacaat
      541 cctctttctc ttttgtttta catgaagggt ctggcagcca aagcaatcac tcaaagttca
      601 aaccttatca ttttttgctt tgttcctctt ggccttggtt ttgtacatca gctttgaaaa
      661 taccatccca gggttaatgc tggggttaat ttataactaa gagtgctcta gttttgcaat
      721 acaggacatg ctataaaaat ggaaagatac cggtgctagc aagcttggat ccaccatggt
      781 gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga
      841 cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa
      901 gctgaccctg aagttcatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt
      961 gaccaccctg acctacggcg tgcagtgctt cagccgctac cccgaccaca tgaagcagca
     1021 cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa
     1081 ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa
     1141 ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct
     1201 ggagtacaac tacaacagcc acaacgtcta tatcatggcc gacaagcaga agaacggcat
     1261 caaggtgaac ttcaagatcc gccacaacat cgaggacggc agcgtgcagc tcgccgacca
     1321 ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct
     1381 gagcacccag tccgccctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct
     1441 ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca agtaagcggc
     1501 tgcacacgtc aatcactagt gaattcgcgg ccgctcgagg tcgacaatca acctctggat
     1561 tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt
     1621 ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc
     1681 tcctccttgt ataaatcctg gttagttctt gccacggcgg aactcatcgc cgcctgcctt
     1741 gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt ggagctcgct
     1801 gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc
     1861 cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg
     1921 catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca
     1981 agggggagga ttgggaagac aatagcaggc atgctgggga gagatcgatc tgaggaaccc
     2041 ctagtgatgg agttggccac tccctctctg cgcgctcgct cgctcactga ggccgggcga
     2101 ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct cagtgagcga gcgagcgcgc
     2161 agctgcctgc aggacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     2221 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     2281 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccccccc
     2341 cccccccggc gattctcttg tttgctccag actctcaggc aatgacctga tagcctttgt
     2401 agagacctct caaaaatagc taccctctcc ggcatgaatt tatcagctag aacggttgaa
     2461 tatcatattg atggtgattt gactgtctcc ggcctttctc acccgtttga atctttacct
     2521 acacattact caggcattgc atttaaaata tatgagggtt ctaaaaattt ttatccttgc
     2581 gttgaaataa aggcttctcc cgcaaaagta ttacagggtc ataatgtttt tggtacaacc
     2641 gatttagctt tatgctctga ggctttattg cttaattttg ctaattcttt gccttgcctg
     2701 tatgatttat tggatgttgg aatcgcctga tgcggtattt tctccttacg catctgtgcg
     2761 gtatttcaca ccgcatatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa
     2821 gccagccccg acacccgcca acactatggt gcactctcag tacaatctgc tctgatgccg
     2881 catagttaag ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc
     2941 tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga
     3001 ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt
     3061 tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa
     3121 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     3181 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
     3241 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     3301 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
     3361 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
     3421 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg
     3481 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
     3541 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     3601 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
     3661 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     3721 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa
     3781 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
     3841 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     3901 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
     3961 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     4021 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     4081 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
     4141 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     4201 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     4261 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     4321 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     4381 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
     4441 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     4501 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     4561 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     4621 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     4681 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     4741 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     4801 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     4861 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
     4921 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc
     4981 gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa
     5041 tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg cgtaatagcg
     5101 aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc gaatggcgat
     5161 tccgttgcaa tggctggcgg taatattgtt ctggatatta ccagcaaggc cgatagtttg
     5221 agttcttcta ctcaggcaag tgatgttatt actaatcaaa gaagtattgc gacaacggtt
     5281 aatttgcgtg atggacagac tcttttactc ggtggcctca ctgattataa aaacacttct
     5341 caggattctg gcgtaccgtt cctgtctaaa atccctttaa tcggcctcct gtttagctcc
     5401 cgctctgatt ctaacgagga aagcacgtta tacgtgctcg tcaaagcaac catagtacgc
     5461 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     5521 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     5581 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
     5641 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc
     5701 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     5761 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg
     5821 gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     5881 gaattttaac aaaatattaa cgcttacaat ttaaatattt gcttatacaa tcttcctgtt
     5941 tttggggctt ttctgattat caaccggggt acatatgatt gacatgctag ttttacgatt
     6001 accgttcatc gcc
//
