LOCUS       sequence-55251-e        6650 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 6650)
  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 6650)
  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: aTRE
FEATURES             Location/Qualifiers
     source          1..6650
                     /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      357..366
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(363..365,366..368,369..1082)
                     /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(363..365,366..368,369..1079)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(408..429)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(669..688)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1016..1037
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    1095..1662
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    1112..1664
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1256..1273)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1483..1502
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             1676..3175
                     /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(3674..3693)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3739..3794
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3793..3812)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4153..4169)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4153..4169)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4166..4188)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4177..4193
                     /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(4201..4207,4208..4225,4226..4231))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4246..4267
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4326..4521
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4372..4507
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    4527..4661
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4564..4583)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4618..4637
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4729..4746)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4900..5488)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4980..4999)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5659..6450,6451..6519))
                     /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     6282..6301
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6520..6624)
                     /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 cgagttattg aggtcagctg
      241 aggtcaagtt acggatccgt aacttgacct cagaattcta tataatggaa gcttgaccgg
      301 agccgttagt tgcccgtcgg acgcgatttg cgaggcgctt tatgcgcgca ccggtcgcca
      361 ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg
      421 acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct
      481 acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca
      541 ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc gaccacatga
      601 agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct
      661 tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc
      721 tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc
      781 acaagctgga gtacaactac aacagccaca acgtctatat catggccgac aagcagaaga
      841 acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc gtgcagctcg
      901 ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc
      961 actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc gatcacatgg
     1021 tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagt
     1081 aaagcggcca attctccctc ccccccccct aacgttactg gccgaagccg cttggaataa
     1141 ggccggtgtg cgtttgtcta tatgttattt tccaccatat tgccgtcttt tggcaatgtg
     1201 agggcccgga aacctggccc tgtcttcttg acgagcattc ctaggggtct ttcccctctc
     1261 gccaaaggaa tgcaaggtct gttgaatgtc gtgaaggaag cagttcctct ggaagcttct
     1321 tgaagacaaa caacgtctgt agcgaccctt tgcaggcagc ggaacccccc acctggcgac
     1381 aggtgcctct gcggccaaaa gccacgtgta taagatacac ctgcaaaggc ggcacaaccc
     1441 cagtgccacg ttgtgagttg gatagttgtg gaaagagtca aatggctctc ctcaagcgta
     1501 ttcaacaagg ggctgaagga tgcccagaag gtaccccatt gtatgggatc tgatctgggg
     1561 cctcggtgca catgctttac gtgtgtttag tcgaggttaa aaaaacgtct aggccccccg
     1621 aaccacgggg acgtggtttt cctttgaaaa acacgatgat aatatggggt cgaggctgct
     1681 gctgctgctg ctgctgggcc tgaggctaca gctctccctg ggcatcatcc tagttgagga
     1741 ggagaacccg gacttctgga accgcgaggc agccgaggcc ctgggtgccg ccaagaagct
     1801 gcagcctgca cagacagccg ccaagaacct catcatcttc ctgggcgatg ggatgggggt
     1861 gtctacggtg acagctgcca ggatcctaaa agggcagaag aaggacaaac tggggcctga
     1921 gatacccctg gccatggacc gcttcccata tgtggctctg tccaagacat acaatgtaga
     1981 caaacatgtg ccagacagtg gagccacagc cacggcctac ctgtgcgggg tcaagggcaa
     2041 cttccagacc attggcttga gtgcagccgc ccgctttaac cagtgcaaca cgacacgcgg
     2101 caacgaggtc atctccgtga tgaatcgggc caagaaagca gggaagtcag tgggagtggt
     2161 aaccaccaca cgagtgcagc acgcctcgcc agccggcacc tacgcccaca cggtgaaccg
     2221 caactggtac tcggacgccg acgtgcctgc ctcggcccgc caggaggggt gccaggacat
     2281 cgctacgcag ctcatctcca acatggacat tgatgtgatc ctaggtggag gccgaaagta
     2341 catgtttcgc atgggaaccc cagaccctga gtacccagat gactacagcc aaggtgggac
     2401 caggctggac gggaagaatc tggtgcagga atggctcggc gaacgccagg gtgcccggta
     2461 cgtgtggaac cgcactgagc tcatgcaggc ttccctggac ccgtctgtga cccatctcat
     2521 gggtctcttt gagcctggag acatgaaata cgagatccac cgagactcca cactggaccc
     2581 ctccctgatg gagatgacag aggctgccct gcgcctgctg agcagacacc cccgcggctt
     2641 cttcctcttc gtggagggtg gtcgcatcga ccatggtcat catgaaagca gggcttaccg
     2701 ggcactgact gagacgatca tgttcgacga cgccattgag agggcgggcc agctcaccag
     2761 cgaggaggac acgctgagcc tcgtcactgc cgaccactcc cacgtcttct ccttcggagg
     2821 ctaccccctg cgagggagct ccttcatcgg gctggccgct ggcaaggccc gggacaggaa
     2881 ggcctacacg gtcctcctat acggaaacgg tccaggctat gtgctcaagg acggcgcccg
     2941 gccggatgtt accgagagcg agagcgggag ccccgagtat cggcagcagt cagcagtgcc
     3001 cctggacgaa gagacccacg caggcgagga cgtggcggtg ttcgcgcgcg gcccgcaggc
     3061 gcacctggtt cacggcgtgc aggagcagac cttcatagcg cacgtcatgg ccttcgccgc
     3121 ctgcctggag ccctacaccg cctgcgacct ggcgcccccc gccggcacca ccgacgccgc
     3181 gcacccgggg cggtccgtgg tccccgcgtt gcttcctctg ctggccggga ccctgctgct
     3241 gctggagacg gccactgctc cctgagtgtc ccgtccctgg ggctcctgct tccccatccc
     3301 ggagttctcc tgctccccac ctcctgtcgt cctgcctggc ctccagcccg agtcgtcatc
     3361 cccggagtcc ctatacagag gtcctgccat ggaaccttcc cctccccgtg cgctctgggg
     3421 actgagccca tgacaccaaa cctgcccctt ggctgctctc ggactcccta ccccaacccc
     3481 agggactgca ggttgtgccc tgtggctgcc tgcaccccag gaaaggaggg ggctcaggcc
     3541 atccagccac cacctacagc ccagtgggta cggtcgagga tatcgcggcc ggccgcactc
     3601 ctcaggtgca ggctgcctat cagaaggtgg tggctggtgt ggccaatgcc ctggctcaca
     3661 aataccactg agatcttttt ccctctgcca aaaattatgg ggacatcatg aagccccttg
     3721 agcatctgac ttctggctaa taaaggaaat ttattttcat tgcaatagtg tgttggaatt
     3781 ttttgtgtct ctcactcgga aggacatatg ggagggcaaa tcatttaaaa catcagaatg
     3841 agtatttggt ttagagtttg gcaacatatg ccatatgctg gctgccatga acaaaggtgg
     3901 ctataaagag gtcatcagta tatgaaacag ccccctgctg tccattcctt attccataga
     3961 aaagccttga cttgaggtta gatttttttt atattttgtt ttgtgttatt tttttcttta
     4021 acatccctaa aattttcctt acatgtttta ctagccagat ttttcctcct ctcctgacta
     4081 ctcccagtca tagctgtccc tcttctctta tgaagatccc tcgacctgca gcccaagctt
     4141 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     4201 caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     4261 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagcg
     4321 gatccgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc
     4381 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
     4441 gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt
     4501 ggaggcctag gcttttgcaa aaagctaact tgtttattgc agcttataat ggttacaaat
     4561 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     4621 gtttgtccaa actcatcaat gtatcttatc atgtctggat ccgctgcatt aatgaatcgg
     4681 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     4741 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     4801 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     4861 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     4921 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     4981 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     5041 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcaatgctc
     5101 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     5161 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     5221 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     5281 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     5341 gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     5401 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     5461 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     5521 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     5581 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     5641 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     5701 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     5761 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     5821 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     5881 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     5941 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     6001 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     6061 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     6121 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     6181 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     6241 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     6301 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     6361 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     6421 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     6481 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     6541 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     6601 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
//
