LOCUS       Exported                9235 bp DNA     circular SYN 06-NOV-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    SFG.wtCNa_opt.IRES.eGFP
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9235)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..9235
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     source          1879..3441
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     source          3497..4058
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    651..1008
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1082..1489
                     /codon_start=1
                     /label=truncated gag
                     /translation="VTTPLSLTLGHWKDVERIAHNQSVDVKKRRWVTFCSAEWPTFNVG
                     WPRDGTFNRDLITQVKIKVFSPGPHGHPDQVPYIVTWEALAFDPPPWVKPFVHPKPPPP
                     LPPSAPSLPLEPPRSTPPRSSLYPALTPSLGA"
     misc_feature    1499..1873
                     /label=pol region
                     /note="Moloney murine leukemia virus (MMLV) pol region 
                     containing the splice acceptor site"
     CDS             1879..3441
                     /gene="PPP3CA"
                     /label=Protein phosphatase 3 catalytic subunit alpha
                     /note="Protein phosphatase 3 catalytic subunit alpha from
                     Homo sapiens. Accession#: Q08209"
     misc_feature    3497..4058
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     CDS             4060..4779
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     LTR             5045..5514
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(6210..6226)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        6700..6804
                     /gene="bla"
                     /label=AmpR promoter
     CDS             6805..7665
                     /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"
     rep_origin      7836..8424
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    8712..8733
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        8748..8778
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8786..8802
                     /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)."
     primer_bind     8810..8826
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     LTR             join(9234..9235,1..588)
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
ORIGIN
        1 agaccccacc tgtaggtttg gcaagctagc ttaagtaacg ccattttgca aggcatggaa
       61 aaatacataa ctgagaatag aaaagttcag atcaaggtca ggaacagatg gaacagctga
      121 atatgggcca aacaggatat ctgtggtaag cagttcctgc cccggctcag ggccaagaac
      181 agatggaaca gctgaatatg ggccaaacag gatatctgtg gtaagcagtt cctgccccgg
      241 ctcagggcca agaacagatg gtccccagat gcggtccagc cctcagcagt ttctagagaa
      301 ccatcagatg tttccagggt gccccaagga cctgaaatga ccctgtgcct tatttgaact
      361 aaccaatcag ttcgcttctc gcttctgttc gcgcgcttct gctccccgag ctcaataaaa
      421 gagcccacaa cccctcactc ggcgcgccag tcctccgatt gactgagtcg cccgggtacc
      481 cgtgtatcca ataaaccctc ttgcagttgc atccgacttg tggtctcgct gttccttggg
      541 agggtctcct ctgagtgatt gactacccgt cagcgggggt ctttcatttg ggggctcgtc
      601 cgggatcggg agacccctgc ccagggacca ccgacccacc accgggaggt aagctggcca
      661 gcaacttatc tgtgtctgtc cgattgtcta gtgtctatga ctgattttat gcgcctgcgt
      721 cggtactagt tagctaacta gctctgtatc tggcggaccc gtggtggaac tgacgagttc
      781 ggaacacccg gccgcaaccc tgggagacgt cccagggact tcgggggccg tttttgtggc
      841 ccgacctgag tcctaaaatc ccgatcgttt aggactcttt ggtgcacccc ccttagagga
      901 gggatatgtg gttctggtag gagacgagaa cctaaaacag ttcccgcctc cgtctgaatt
      961 tttgctttcg gtttgggacc gaagccgcgc cgcgcgtctt gtctgctgca gcatcgttct
     1021 gtgttgtctc tgtctgactg tgtttctgta tttgtctgaa aatatgggcc cgggctagcc
     1081 tgttaccact cccttaagtt tgaccttagg tcactggaaa gatgtcgagc ggatcgctca
     1141 caaccagtcg gtagatgtca agaagagacg ttgggttacc ttctgctctg cagaatggcc
     1201 aacctttaac gtcggatggc cgcgagacgg cacctttaac cgagacctca tcacccaggt
     1261 taagatcaag gtcttttcac ctggcccgca tggacaccca gaccaggtcc cctacatcgt
     1321 gacctgggaa gccttggctt ttgacccccc tccctgggtc aagccctttg tacaccctaa
     1381 gcctccgcct cctcttcctc catccgcccc gtctctcccc cttgaacctc ctcgttcgac
     1441 cccgcctcga tcctcccttt atccagccct cactccttct ctaggcgccc ccatatggcc
     1501 atatgagatc ttatatgggg cacccccgcc ccttgtaaac ttccctgacc ctgacatgac
     1561 aagagttact aacagcccct ctctccaagc tcacttacag gctctctact tagtccagca
     1621 cgaagtctgg agacctctgg cggcagccta ccaagaacaa ctggaccgac cggtggtacc
     1681 tcacccttac cgagtcggcg acacagtgtg ggtccgccga caccagacta agaacctaga
     1741 acctcgctgg aaaggacctt acacagtcct gctgaccacc cccaccgccc tcaaagtaga
     1801 cggcatcgca gcttggatac acgccgccca cgtgaaggct gccgaccccg ggggtggacc
     1861 atcctctaga ctgccaccat gagcgagccc aaggccatcg accccaagct gtccaccacc
     1921 gacagagtgg tgaaggccgt gccctttccc ccctcccacc gcctgaccgc caaggaggtg
     1981 ttcgacaacg atggcaagcc cagagtggac atcctgaaag cccacctgat gaaggagggc
     2041 agactggagg agagcgtggc cctgcgcatc atcaccgagg gcgccagcat cctgcgccag
     2101 gagaagaacc tgctggatat cgacgcccct gtgaccgtgt gcggcgacat ccacggacag
     2161 tttttcgacc tgatgaagct gttcgaggtg ggcggctctc ccgccaacac cagatacctg
     2221 ttcctgggcg actacgtgga cagaggctac ttctctatcg agtgcgtgct gtacctgtgg
     2281 gccctgaaga tcctgtaccc caagaccctg ttcctgctga gaggcaacca cgagtgccgc
     2341 cacctgaccg agtactttac cttcaagcag gagtgcaaga tcaagtacag cgagcgcgtg
     2401 tacgatgcct gcatggacgc cttcgactgt ctgcccctgg ccgctctgat gaaccagcag
     2461 ttcctgtgtg tgcacggagg cctgagcccc gagatcaaca ccctggacga catccggaag
     2521 ctggaccgct tcaaggagcc ccctgcctat ggccccatgt gcgacatcct gtggagcgac
     2581 cccctggagg actttggcaa cgagaagacc caggagcact ttacccacaa caccgtgaga
     2641 ggctgcagct acttctactc ctaccctgct gtgtgcgagt tcctgcagca caacaacctg
     2701 ctgagcatcc tgagggccca cgaggcccag gatgccggct accgcatgta ccgcaagtcc
     2761 cagaccaccg gcttcccctc cctgatcacc atcttctctg cccccaacta cctggacgtg
     2821 tacaacaaca aagccgctgt gctgaagtac gagaacaacg tgatgaacat ccggcagttc
     2881 aactgctccc cccaccccta ctggctgccc aacttcatgg acgtgttcac ctggtccctg
     2941 ccctttgtgg gcgagaaggt gaccgagatg ctggtgaacg tgctgaacat ctgctccgac
     3001 gatgagctgg gcagcgagga ggacggcttc gatggcgcca ccgctgccgc cagaaaggag
     3061 gtgatccgca ataagatcag agccatcggc aagatggcca gagtgttctc cgtgctgcgc
     3121 gaggagagcg agtccgtgct gaccctgaag ggcctgaccc ccaccggcat gctgccctcc
     3181 ggcgtgctga gcggaggcaa gcagaccctg cagagcgcca cagtggaggc catcgaggcc
     3241 gatgaggcca tcaagggctt ctccccccag cacaagatca ccagcttcga ggaggccaag
     3301 ggcctggacc ggatcaacga gcggatgcca cccagacggg atgccatgcc cagcgatgcc
     3361 aacctgaact ccatcaacaa ggccctgacc agcgagacca acggcaccga cagcaacggc
     3421 agcaactcct ccaacatcca gtgaacgcgt acgtcgcgac cgcggacatg tacagagctc
     3481 gagcgggatc aattccgccc cccccctaac gttactggcc gaagccgctt ggaataaggc
     3541 cggtgtgcgt ttgtctatat gttattttcc accatattgc cgtcttttgg caatgtgagg
     3601 gcccggaaac ctggccctgt cttcttgacg agcattccta ggggtctttc ccctctcgcc
     3661 aaaggaatgc aaggtctgtt gaatgtcgtg aaggaagcag ttcctctgga agcttcttga
     3721 agacaaacaa cgtctgtagc gaccctttgc aggcagcgga accccccacc tggcgacagg
     3781 tgcctctgcg gccaaaagcc acgtgtataa gatacacctg caaaggcggc acaaccccag
     3841 tgccacgttg tgagttggat agttgtggaa agagtcaaat ggctctcctc aagcgtattc
     3901 aacaaggggc tgaaggatgc ccagaaggta ccccattgta tgggatctga tctggggcct
     3961 cggtgcacat gctttacatg tgtttagtcg aggttaaaaa acgtctaggc cccccgaacc
     4021 acggggacgt ggttttcctt tgaaaaacac gataataata tggtgagcaa gggcgaggag
     4081 ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag
     4141 ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagttc
     4201 atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgacctac
     4261 ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc
     4321 gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac
     4381 aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag
     4441 ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac
     4501 agccacaacg tctatatcat ggccgacaag cagaagaacg gcatcaaggt gaacttcaag
     4561 atccgccaca acatcgagga cggcagcgtg cagctcgccg accactacca gcagaacacc
     4621 cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcac ccagtccgcc
     4681 ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc
     4741 gccgggatca ctctcggcat ggacgagctg tacaagtaaa gcggattagt ccaatttgtt
     4801 aaagacagga tatcagtggt ccaggctcta gttttgactc aacaatatca ccagctgaag
     4861 cctatagagt acgagccata gataaaataa aagattttat ttagtctcca gaaaaagggg
     4921 ggaatgaaag accccacctg taggtttggc aagctagctt aagtaacgcc attttgcaag
     4981 gcatggaaaa atacataact gagaatagag aagttcagat caaggtcagg aacagatgga
     5041 acagctgaat atgggccaaa caggatatct gtggtaagca gttcctgccc cggctcaggg
     5101 ccaagaacag atggaacagc tgaatatggg ccaaacagga tatctgtggt aagcagttcc
     5161 tgccccggct cagggccaag aacagatggt ccccagatgc ggtccagccc tcagcagttt
     5221 ctagagaacc atcagatgtt tccagggtgc cccaaggacc tgaaatgacc ctgtgcctta
     5281 tttgaactaa ccaatcagtt cgcttctcgc ttctgttcgc gcgcttctgc tccccgagct
     5341 caataaaaga gcccacaacc cctcactcgg ggcgccagtc ctccgattga ctgagtcgcc
     5401 cgggtacccg tgtatccaat aaaccctctt gcagttgcat ccgacttgtg gtctcgctgt
     5461 tccttgggag ggtctcctct gagtgattga ctacccgtca gcgggggtct ttcacatgca
     5521 gcatgtatca aaattaattt ggtttttttt cttaagtatt tacattaaat ggccatagta
     5581 cttaaagtta cattggcttc cttgaaataa acatggagta ttcagaatgt gtcataaata
     5641 tttctaattt taagatagta tctccattgg ctttctactt tttcttttat ttttttttgt
     5701 cctctgtctt ccatttgttg ttgttgttgt ttgtttgttt gtttgttggt tggttggtta
     5761 attttttttt aaagatccta cactatagtt caagctagac tattagctac tctgtaaccc
     5821 agggtgacct tgaagtcatg ggtagcctgc tgttttagcc ttcccacatc taagattaca
     5881 ggtatgagct atcatttttg gtatattgat tgattgattg attgatgtgt gtgtgtgtga
     5941 ttgtgtttgt gtgtgtgatt gtgtatatgt gtgtatggtt gtgtgtgatt gtgtgtatgt
     6001 atgtttgtgt gtgattgtgt gtgtgtgatt gtgcatgtgt gtgtgtgtga ttgtgtttat
     6061 gtgtatgatt gtgtgtgtgt gtgtgtgtgt gtgtgtgtgt gtgtgtgtgt gtgtgttgtg
     6121 tatatatatt tatggtagtg agaggcaacg ctccggctca ggtgtcaggt tggtttttga
     6181 gacagagtct ttcacttagc ttggaattca ctggccgtcg ttttacaacg tcgtgactgg
     6241 gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt cgccagctgg
     6301 cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc
     6361 gaatggcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata
     6421 tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagcc ccgacacccg
     6481 ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa
     6541 gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc
     6601 gcgagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg
     6661 gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     6721 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     6781 caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc
     6841 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa
     6901 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt
     6961 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt
     7021 ctgctatgtg gcgcggtatt atcccgtatt gatgccgggc aagagcaact cggtcgccgc
     7081 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg
     7141 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg
     7201 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac
     7261 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca
     7321 aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta
     7381 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat
     7441 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa
     7501 tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag
     7561 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat
     7621 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt
     7681 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     7741 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     7801 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     7861 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     7921 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     7981 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     8041 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     8101 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     8161 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     8221 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     8281 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     8341 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     8401 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     8461 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     8521 cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc
     8581 gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt
     8641 tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag
     8701 cgcaacgcaa ttaatgtgag ttagctcact cattaggcac cccaggcttt acactttatg
     8761 cttccggctc gtatgttgtg tggaattgtg agcggataac aatttcacac aggaaacagc
     8821 tatgaccatg attacgccaa gctttgctct taggagtttc ctaatacatc ccaaactcaa
     8881 atatataaag catttgactt gttctatgcc ctagggggcg gggggaagct aagccagctt
     8941 tttttaacat ttaaaatgtt aattccattt taaatgcaca gatgttttta tttcataagg
     9001 gtttcaatgt gcatgaatgc tgcaatattc ctgttaccaa agctagtata aataaaaata
     9061 gataaacgtg gaaattactt agagtttctg tcattaacgt ttccttcctc agttgacaac
     9121 ataaatgcgc tgctgagaag ccagtttgca tctgtcagga tcaatttccc attatgccag
     9181 tcatattaat tactagtcaa ttagttgatt tttatttttg acatatacat gtgaa
//
