LOCUS       sequence-55252-e        6630 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 6630)
  AUTHORS   Billings NA, Emerson MM, Cepko CL
  TITLE     Analysis of thyroid response element activity during retinal 
            development.
  JOURNAL   PLoS One. 2010  . 5(10):e13739.
  PUBMED    21060789
REFERENCE   2  (bases 1 to 6630)
  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: "PLoS One. 
            2010  . 5(10):e13739."
COMMENT     Sequence Label: rME
FEATURES             Location/Qualifiers
     source          1..6630
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    46..94
                     /note="synthetic polyadenylation signal"
     misc_feature    108..199
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     148..167
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     regulatory      337..346
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(343..345,346..348,349..1062)
                     /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(343..345,346..348,349..1059)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(388..409)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(649..668)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     996..1017
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    1075..1642
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    1092..1644
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1236..1253)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1463..1482
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             1656..3155
                     /codon_start=1
                     /product="secreted alkaline phosphatase from human 
                     placenta"
                     /label=SEAP
                     /note="The natural enzyme has a C-terminal transmembrane 
                     sequence that is replaced by a GPI anchor."
                     /translation="LLLLLLLGLRLQLSLGIILVEEENPDFWNREAAEALGAAKKLQPA
                     QTAAKNLIIFLGDGMGVSTVTAARILKGQKKDKLGPEIPLAMDRFPYVALSKTYNVDKH
                     VPDSGATATAYLCGVKGNFQTIGLSAAARFNQCNTTRGNEVISVMNRAKKAGKSVGVVT
                     TTRVQHASPAGTYAHTVNRNWYSDADVPASARQEGCQDIATQLISNMDIDVILGGGRKY
                     MFRMGTPDPEYPDDYSQGGTRLDGKNLVQEWLGERQGARYVWNRTELMQASLDPSVTHL
                     MGLFEPGDMKYEIHRDSTLDPSLMEMTEAALRLLSRHPRGFFLFVEGGRIDHGHHESRA
                     YRALTETIMFDDAIERAGQLTSEEDTLSLVTADHSHVFSFGGYPLRGSSFIGLAAGKAR
                     DRKAYTVLLYGNGPGYVLKDGARPDVTESESGSPEYRQQSAVPLDEETHAGEDVAVFAR
                     GPQAHLVHGVQEQTFIAHVMAFAACLEPYTACDLAPPAGTTD"
     primer_bind     complement(3654..3673)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3719..3774
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3773..3792)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4133..4149)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4133..4149)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4146..4168)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4157..4173
                     /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)."
     promoter        complement(join(4181..4187,4188..4205,4206..4211))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4226..4247
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4306..4501
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4352..4487
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    4507..4641
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4544..4563)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4598..4617
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4709..4726)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4880..5468)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4960..4979)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5639..6430,6431..6499))
                     /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     6262..6281
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6500..6604)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgagaagt aatattaagg tacgggaggt acttggagcg gccgcaataa aatatcttta
       61 ttttcattac atctgtgtgt tggttttttg tgtgaatcga tagtactaac atacgctctc
      121 catcaaaaca aaacgaaaca aaacaaacta gcaaaatagg ctgtccccag tgcaagtgca
      181 ggtgccagaa catttctcta tcgatagtcg accaattgct cgagaggacg ttggggttag
      241 gggaggacag tggaattcta tataatggaa gcttgaccgg agccgttagt tgcccgtcgg
      301 acgcgatttg cgaggcgctt tatgcgcgca ccggtcgcca ccatggtgag caagggcgag
      361 gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac
      421 aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag
      481 ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgacc
      541 tacggcgtgc agtgcttcag ccgctacccc gaccacatga agcagcacga cttcttcaag
      601 tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac
      661 tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg
      721 aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac
      781 aacagccaca acgtctatat catggccgac aagcagaaga acggcatcaa ggtgaacttc
      841 aagatccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac
      901 acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag cacccagtcc
      961 gccctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc
     1021 gccgccggga tcactctcgg catggacgag ctgtacaagt aaagcggcca attctccctc
     1081 ccccccccct aacgttactg gccgaagccg cttggaataa ggccggtgtg cgtttgtcta
     1141 tatgttattt tccaccatat tgccgtcttt tggcaatgtg agggcccgga aacctggccc
     1201 tgtcttcttg acgagcattc ctaggggtct ttcccctctc gccaaaggaa tgcaaggtct
     1261 gttgaatgtc gtgaaggaag cagttcctct ggaagcttct tgaagacaaa caacgtctgt
     1321 agcgaccctt tgcaggcagc ggaacccccc acctggcgac aggtgcctct gcggccaaaa
     1381 gccacgtgta taagatacac ctgcaaaggc ggcacaaccc cagtgccacg ttgtgagttg
     1441 gatagttgtg gaaagagtca aatggctctc ctcaagcgta ttcaacaagg ggctgaagga
     1501 tgcccagaag gtaccccatt gtatgggatc tgatctgggg cctcggtgca catgctttac
     1561 gtgtgtttag tcgaggttaa aaaaacgtct aggccccccg aaccacgggg acgtggtttt
     1621 cctttgaaaa acacgatgat aatatggggt cgaggctgct gctgctgctg ctgctgggcc
     1681 tgaggctaca gctctccctg ggcatcatcc tagttgagga ggagaacccg gacttctgga
     1741 accgcgaggc agccgaggcc ctgggtgccg ccaagaagct gcagcctgca cagacagccg
     1801 ccaagaacct catcatcttc ctgggcgatg ggatgggggt gtctacggtg acagctgcca
     1861 ggatcctaaa agggcagaag aaggacaaac tggggcctga gatacccctg gccatggacc
     1921 gcttcccata tgtggctctg tccaagacat acaatgtaga caaacatgtg ccagacagtg
     1981 gagccacagc cacggcctac ctgtgcgggg tcaagggcaa cttccagacc attggcttga
     2041 gtgcagccgc ccgctttaac cagtgcaaca cgacacgcgg caacgaggtc atctccgtga
     2101 tgaatcgggc caagaaagca gggaagtcag tgggagtggt aaccaccaca cgagtgcagc
     2161 acgcctcgcc agccggcacc tacgcccaca cggtgaaccg caactggtac tcggacgccg
     2221 acgtgcctgc ctcggcccgc caggaggggt gccaggacat cgctacgcag ctcatctcca
     2281 acatggacat tgatgtgatc ctaggtggag gccgaaagta catgtttcgc atgggaaccc
     2341 cagaccctga gtacccagat gactacagcc aaggtgggac caggctggac gggaagaatc
     2401 tggtgcagga atggctcggc gaacgccagg gtgcccggta cgtgtggaac cgcactgagc
     2461 tcatgcaggc ttccctggac ccgtctgtga cccatctcat gggtctcttt gagcctggag
     2521 acatgaaata cgagatccac cgagactcca cactggaccc ctccctgatg gagatgacag
     2581 aggctgccct gcgcctgctg agcagacacc cccgcggctt cttcctcttc gtggagggtg
     2641 gtcgcatcga ccatggtcat catgaaagca gggcttaccg ggcactgact gagacgatca
     2701 tgttcgacga cgccattgag agggcgggcc agctcaccag cgaggaggac acgctgagcc
     2761 tcgtcactgc cgaccactcc cacgtcttct ccttcggagg ctaccccctg cgagggagct
     2821 ccttcatcgg gctggccgct ggcaaggccc gggacaggaa ggcctacacg gtcctcctat
     2881 acggaaacgg tccaggctat gtgctcaagg acggcgcccg gccggatgtt accgagagcg
     2941 agagcgggag ccccgagtat cggcagcagt cagcagtgcc cctggacgaa gagacccacg
     3001 caggcgagga cgtggcggtg ttcgcgcgcg gcccgcaggc gcacctggtt cacggcgtgc
     3061 aggagcagac cttcatagcg cacgtcatgg ccttcgccgc ctgcctggag ccctacaccg
     3121 cctgcgacct ggcgcccccc gccggcacca ccgacgccgc gcacccgggg cggtccgtgg
     3181 tccccgcgtt gcttcctctg ctggccggga ccctgctgct gctggagacg gccactgctc
     3241 cctgagtgtc ccgtccctgg ggctcctgct tccccatccc ggagttctcc tgctccccac
     3301 ctcctgtcgt cctgcctggc ctccagcccg agtcgtcatc cccggagtcc ctatacagag
     3361 gtcctgccat ggaaccttcc cctccccgtg cgctctgggg actgagccca tgacaccaaa
     3421 cctgcccctt ggctgctctc ggactcccta ccccaacccc agggactgca ggttgtgccc
     3481 tgtggctgcc tgcaccccag gaaaggaggg ggctcaggcc atccagccac cacctacagc
     3541 ccagtgggta cggtcgagga tatcgcggcc ggccgcactc ctcaggtgca ggctgcctat
     3601 cagaaggtgg tggctggtgt ggccaatgcc ctggctcaca aataccactg agatcttttt
     3661 ccctctgcca aaaattatgg ggacatcatg aagccccttg agcatctgac ttctggctaa
     3721 taaaggaaat ttattttcat tgcaatagtg tgttggaatt ttttgtgtct ctcactcgga
     3781 aggacatatg ggagggcaaa tcatttaaaa catcagaatg agtatttggt ttagagtttg
     3841 gcaacatatg ccatatgctg gctgccatga acaaaggtgg ctataaagag gtcatcagta
     3901 tatgaaacag ccccctgctg tccattcctt attccataga aaagccttga cttgaggtta
     3961 gatttttttt atattttgtt ttgtgttatt tttttcttta acatccctaa aattttcctt
     4021 acatgtttta ctagccagat ttttcctcct ctcctgacta ctcccagtca tagctgtccc
     4081 tcttctctta tgaagatccc tcgacctgca gcccaagctt ggcgtaatca tggtcatagc
     4141 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
     4201 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
     4261 cactgcccgc tttccagtcg ggaaacctgt cgtgccagcg gatccgcatc tcaattagtc
     4321 agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc
     4381 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
     4441 ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa
     4501 aaagctaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     4561 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     4621 gtatcttatc atgtctggat ccgctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     4681 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     4741 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     4801 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     4861 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     4921 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     4981 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     5041 cctttctccc ttcgggaagc gtggcgcttt ctcaatgctc acgctgtagg tatctcagtt
     5101 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     5161 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     5221 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     5281 agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg
     5341 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     5401 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     5461 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     5521 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     5581 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     5641 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     5701 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     5761 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     5821 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     5881 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     5941 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     6001 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     6061 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     6121 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     6181 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     6241 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     6301 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     6361 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     6421 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     6481 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     6541 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     6601 cgcgcacatt tccccgaaaa gtgccacctg
//
