LOCUS       sequence-252277-        7749 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7749)
  AUTHORS   Seoane J, Le HV, Shen L, Anderson SA, Massague J
  TITLE     Integration of Smad and forkhead pathways in the control of 
            neuroepithelial and glioblastoma cell proliferation.
  JOURNAL   Cell. 2004 Apr 16. 117(2):211-23.
  PUBMED    15084259
REFERENCE   2  (bases 1 to 7749)
  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: "Cell. 2004 
            Apr 16. 117(2):211-23."
COMMENT     Sequence Label: IRES-GFP FoxG1
FEATURES             Location/Qualifiers
     source          1..7749
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             766..795
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             769..798
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     misc_feature    join(3048..3622,3623..3625,3626..3634)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3215..3232)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3442..3461
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(3635..3637,3638..3640,3641..4354)
                     /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(3635..3637,3638..3640,3641..4351)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3680..3701)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3941..3960)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4288..4309
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    4477..4598
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4514..4533)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4568..4587
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4605..5060)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4742..4763)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4954..4973
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5087..5191
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5193..5550
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5193..5384
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5216..5236)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5401..5536
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5585..6379
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             5585..6379
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5639..5658)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6249..6268
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6567..6586)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6611..6658
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6987..7575
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7476..7495
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      541 ccggactcag atctcgagct caagcttcga attcgtcgag agcgggctcc cgcgccgccg
      601 ccgccgtttc cccccgacga ctgggtgatg ctggacatgg gagataggaa agaggtgaaa
      661 atgatcccca agtcctcgtt cagcatcaac agcctggtgc ccgaggcggt ccagaacgac
      721 aaccaccacg cgagccacgg ccaccacaac agccaccacc cccagcacca ccaccaccac
      781 caccaccatc accaccaccc gccgccgccc gccccgcaac cgccgccgcc gccgcagcag
      841 cagcagccgc cgccgccgcc gcccccggca cggcagcccc cccagacgcg gggcgccccg
      901 gccgccgacg acgacaaggg cccccagcag ctgctgctcc cgccgccgcc accgccacca
      961 ccggccgccg ccctggacgg ggctaaagcg gacgggctgg gcggcaaggg cgagccgggc
     1021 ggcgggccgg gggagctggc gcccgtcggg ccggacgaga aggagaaggg cgccggcgcc
     1081 gggggggagg agaagaaggg ggcgggcgag ggcggcaagg acggggaggg gggcaaggag
     1141 ggcgagaaga agaacggcaa gtacgagaag ccgccgttca gctacaacgc gctcatcatg
     1201 atggccatcc ggcagagccc cgagaagcgg ctcacgctca acggcatcta cgagttcatc
     1261 atgaagaact tcccttacta ccgcgagaac aagcagggct ggcagaactc catccgccac
     1321 aatctgtccc tcaacaagtg cttcgtgaag gtgccgcgcc actacgacga cccgggcaag
     1381 ggcaactact ggatgctgga cccgtcgagc gacgacgtgt tcatcggcgg caccacgggc
     1441 aagctgcggc gccgctccac cacctcgcgg gccaagctgg ccttcaagcg cggtgcgcgc
     1501 ctcacctcca ccggcctcac cttcatggac cgcgccggct ccctctactg gcccatgtcg
     1561 cccttcctgt ccctgcacca cccccgcgcc agcagcactt tgagttacaa cggcaccacg
     1621 tcggcctacc ccagccaccc catgccctac agctccgtgt tgactcagaa ctcgctgggc
     1681 aacaaccact ccttctccac cgccaacggc ctgagcgtgg accggctggt caacggggag
     1741 atcccgtacg ccacgcacca cctcacggcc gccgcgctag ccgcctcggt gccctgcggc
     1801 ctgtcggtgc cctgctctgg gacctactcc ctcaacccct gctccgtcaa cctgctcgcg
     1861 ggccagacca gttacttttt cccccacgtc ccgcacccgt caatgacttc gcagagcagc
     1921 acgtccatga gcgccagggc cgcgtcctcc tccacgtcgc cgcaggcccc ctcgaccctg
     1981 ccctgtgagt ctttaagacc ctctttgcca agttttacga cgggactgtc tgggggactg
     2041 tctgattatt tcacacatca aaatcagggg tcttcttcca accctttaat acattaacat
     2101 ccctgggacc agactgtaag tgaacgtttt acacacattt gcattgtaaa tgataattaa
     2161 aaaaataagt ccaggtattt tttattaagc ccccccctcc catttctgta cgtttgttca
     2221 gtctctaggg ttgtttatta ttctaacaag gtgtggagtg tcagcgaggt gcaatgtggg
     2281 gagaatacat tgtagaatat aaggtttgga agtcaaatta tagtagaatg tgtatctaaa
     2341 tagtgactgc tttgccattt cattcaaacc tgacaagtct atctctaaga gccgccagat
     2401 ttccatgtgt gcagtattat aagttatcat ggaactatat ggtggacgca gaccttgaga
     2461 acaacctaaa ttatggggag aattttaaaa tgttaaactg taatttgtat ttaaaaagca
     2521 ttcgtagtaa aggtgcccaa gaaattattt tggccattta ttgttttgtc cttttcttta
     2581 aagaactgtt tttttttctt ttgtttactt ttagaccaaa gattgggttc tagaaaatgc
     2641 acttggtata ctaagtatta aaacaaacaa aaaggaaagt tgtttcagtt ggcaacactg
     2701 cccattcaat tgaatcagaa ggggacaaaa ttaacgattg ccttcagttt gtgttgtgta
     2761 tattttgatg tatgtggtca ctaacaggtc acttttattt tttctaaatg tagtgaaatg
     2821 ttaataccta ttgtacttat aggtaaacct tgcaaatatg taacctgtgt tgcgcaaatg
     2881 ccgcataaat ttgagtgatt gttaatgttg tcttaaaatt tcttgattgt gatactgtgg
     2941 tcatatgccc gtgtttgtca cttacaaaaa tgtttactat gaacacacag aaataaaaaa
     3001 taggctaaat ccaaaaaaaa aaaaaaaact ctccagcgct ggatccgccc ctctccctcc
     3061 ccccccccta acgttactgg ccgaagccgc ttggaataag gccggtgtgc gtttgtctat
     3121 atgttatttt ccaccatatt gccgtctttt ggcaatgtga gggcccggaa acctggccct
     3181 gtcttcttga cgagcattcc taggggtctt tcccctctcg ccaaaggaat gcaaggtctg
     3241 ttgaatgtcg tgaaggaagc agttcctctg gaagcttctt gaagacaaac aacgtctgta
     3301 gcgacccttt gcaggcagcg gaacccccca cctggcgaca ggtgcctctg cggccaaaag
     3361 ccacgtgtat aagatacacc tgcaaaggcg gcacaacccc agtgccacgt tgtgagttgg
     3421 atagttgtgg aaagagtcaa atggctctcc tcaagcgtat tcaacaaggg gctgaaggat
     3481 gcccagaagg taccccattg tatgggatct gatctggggc ctcggtacac atgctttaca
     3541 tgtgtttagt cgaggttaaa aaaacgtcta ggccccccga accacgggga cgtggttttc
     3601 ctttgaaaaa cacgatgata atatggccac aaccatggtg agcaagggcg aggagctgtt
     3661 caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag
     3721 cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg
     3781 caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctga cctacggcgt
     3841 gcagtgcttc agccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat
     3901 gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac
     3961 ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat
     4021 cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca
     4081 caacgtctat atcatggccg acaagcagaa gaacggcatc aaggtgaact tcaagatccg
     4141 ccacaacatc gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat
     4201 cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agcacccagt ccgccctgag
     4261 caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg
     4321 gatcactctc ggcatggacg agctgtacaa gtaaagcggc cgcgactcta gatcataatc
     4381 agccatacca catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg
     4441 aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat
     4501 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     4561 tctagttgtg gtttgtccaa actcatcaat gtatcttaag gcgtaaattg taagcgttaa
     4621 tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc
     4681 cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt
     4741 tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa
     4801 aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg
     4861 gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg
     4921 acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc
     4981 tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa
     5041 tgcgccgcta cagggcgcgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat
     5101 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     5161 aatgcttcaa taatattgaa aaaggaagag tcctgaggcg gaaagaacca gctgtggaat
     5221 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     5281 atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga
     5341 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
     5401 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
     5461 tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga
     5521 ggcttttttg gaggcctagg cttttgcaaa gatcgatcaa gagacaggat gaggatcgtt
     5581 tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct
     5641 attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct
     5701 gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga
     5761 actgcaagac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc
     5821 tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg
     5881 gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc
     5941 aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca
     6001 tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga
     6061 cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg cgagcatgcc
     6121 cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga
     6181 aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca
     6241 ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg
     6301 cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct
     6361 tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc
     6421 aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga
     6481 atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc
     6541 ttcgcccacc ctagggggag gctaactgaa acacggaagg agacaatacc ggaaggaacc
     6601 cgcgctatga cggcaataaa aagacagaat aaaacgcacg gtgttgggtc gtttgttcat
     6661 aaacgcgggg ttcggtccca gggctggcac tctgtcgata ccccaccgag accccattgg
     6721 ggccaatacg cccgcgtttc ttccttttcc ccaccccacc ccccaagttc gggtgaaggc
     6781 ccagggctcg cagccaacgt cggggcggca ggccctgcca tagcctcagg ttactcatat
     6841 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     6901 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     6961 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     7021 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     7081 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
     7141 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     7201 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     7261 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     7321 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     7381 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     7441 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     7501 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     7561 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     7621 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
     7681 gccatgcatt agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat
     7741 ggagttccg
//
