LOCUS       sequence-55256-e        6631 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 6631)
  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 6631)
  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: cLS-F2
FEATURES             Location/Qualifiers
     source          1..6631
                     /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      338..347
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(344..346,347..349,350..1063)
                     /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(344..346,347..349,350..1060)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(389..410)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(650..669)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     997..1018
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    1076..1643
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    1093..1645
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1237..1254)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1464..1483
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             1657..3156
                     /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(3655..3674)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3720..3775
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3774..3793)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4134..4150)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4134..4150)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4147..4169)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4158..4174
                     /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(4182..4188,4189..4206,4207..4212))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4227..4248
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4307..4502
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4353..4488
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    4508..4642
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4545..4564)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4599..4618
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4710..4727)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4881..5469)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4961..4980)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5640..6431,6432..6500))
                     /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     6263..6282
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6501..6605)
                     /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 accccagctg
      241 aggtcaagtt acggaattct atataatgga agcttgaccg gagccgttag ttgcccgtcg
      301 gacgcgattt gcgaggcgct ttatgcgcgc accggtcgcc accatggtga gcaagggcga
      361 ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca
      421 caagttcagc gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa
      481 gttcatctgc accaccggca agctgcccgt gccctggccc accctcgtga ccaccctgac
      541 ctacggcgtg cagtgcttca gccgctaccc cgaccacatg aagcagcacg acttcttcaa
      601 gtccgccatg cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa
      661 ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct
      721 gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta
      781 caacagccac aacgtctata tcatggccga caagcagaag aacggcatca aggtgaactt
      841 caagatccgc cacaacatcg aggacggcag cgtgcagctc gccgaccact accagcagaa
      901 cacccccatc ggcgacggcc ccgtgctgct gcccgacaac cactacctga gcacccagtc
      961 cgccctgagc aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac
     1021 cgccgccggg atcactctcg gcatggacga gctgtacaag taaagcggcc aattctccct
     1081 cccccccccc taacgttact ggccgaagcc gcttggaata aggccggtgt gcgtttgtct
     1141 atatgttatt ttccaccata ttgccgtctt ttggcaatgt gagggcccgg aaacctggcc
     1201 ctgtcttctt gacgagcatt cctaggggtc tttcccctct cgccaaagga atgcaaggtc
     1261 tgttgaatgt cgtgaaggaa gcagttcctc tggaagcttc ttgaagacaa acaacgtctg
     1321 tagcgaccct ttgcaggcag cggaaccccc cacctggcga caggtgcctc tgcggccaaa
     1381 agccacgtgt ataagataca cctgcaaagg cggcacaacc ccagtgccac gttgtgagtt
     1441 ggatagttgt ggaaagagtc aaatggctct cctcaagcgt attcaacaag gggctgaagg
     1501 atgcccagaa ggtaccccat tgtatgggat ctgatctggg gcctcggtgc acatgcttta
     1561 cgtgtgttta gtcgaggtta aaaaaacgtc taggcccccc gaaccacggg gacgtggttt
     1621 tcctttgaaa aacacgatga taatatgggg tcgaggctgc tgctgctgct gctgctgggc
     1681 ctgaggctac agctctccct gggcatcatc ctagttgagg aggagaaccc ggacttctgg
     1741 aaccgcgagg cagccgaggc cctgggtgcc gccaagaagc tgcagcctgc acagacagcc
     1801 gccaagaacc tcatcatctt cctgggcgat gggatggggg tgtctacggt gacagctgcc
     1861 aggatcctaa aagggcagaa gaaggacaaa ctggggcctg agatacccct ggccatggac
     1921 cgcttcccat atgtggctct gtccaagaca tacaatgtag acaaacatgt gccagacagt
     1981 ggagccacag ccacggccta cctgtgcggg gtcaagggca acttccagac cattggcttg
     2041 agtgcagccg cccgctttaa ccagtgcaac acgacacgcg gcaacgaggt catctccgtg
     2101 atgaatcggg ccaagaaagc agggaagtca gtgggagtgg taaccaccac acgagtgcag
     2161 cacgcctcgc cagccggcac ctacgcccac acggtgaacc gcaactggta ctcggacgcc
     2221 gacgtgcctg cctcggcccg ccaggagggg tgccaggaca tcgctacgca gctcatctcc
     2281 aacatggaca ttgatgtgat cctaggtgga ggccgaaagt acatgtttcg catgggaacc
     2341 ccagaccctg agtacccaga tgactacagc caaggtggga ccaggctgga cgggaagaat
     2401 ctggtgcagg aatggctcgg cgaacgccag ggtgcccggt acgtgtggaa ccgcactgag
     2461 ctcatgcagg cttccctgga cccgtctgtg acccatctca tgggtctctt tgagcctgga
     2521 gacatgaaat acgagatcca ccgagactcc acactggacc cctccctgat ggagatgaca
     2581 gaggctgccc tgcgcctgct gagcagacac ccccgcggct tcttcctctt cgtggagggt
     2641 ggtcgcatcg accatggtca tcatgaaagc agggcttacc gggcactgac tgagacgatc
     2701 atgttcgacg acgccattga gagggcgggc cagctcacca gcgaggagga cacgctgagc
     2761 ctcgtcactg ccgaccactc ccacgtcttc tccttcggag gctaccccct gcgagggagc
     2821 tccttcatcg ggctggccgc tggcaaggcc cgggacagga aggcctacac ggtcctccta
     2881 tacggaaacg gtccaggcta tgtgctcaag gacggcgccc ggccggatgt taccgagagc
     2941 gagagcggga gccccgagta tcggcagcag tcagcagtgc ccctggacga agagacccac
     3001 gcaggcgagg acgtggcggt gttcgcgcgc ggcccgcagg cgcacctggt tcacggcgtg
     3061 caggagcaga ccttcatagc gcacgtcatg gccttcgccg cctgcctgga gccctacacc
     3121 gcctgcgacc tggcgccccc cgccggcacc accgacgccg cgcacccggg gcggtccgtg
     3181 gtccccgcgt tgcttcctct gctggccggg accctgctgc tgctggagac ggccactgct
     3241 ccctgagtgt cccgtccctg gggctcctgc ttccccatcc cggagttctc ctgctcccca
     3301 cctcctgtcg tcctgcctgg cctccagccc gagtcgtcat ccccggagtc cctatacaga
     3361 ggtcctgcca tggaaccttc ccctccccgt gcgctctggg gactgagccc atgacaccaa
     3421 acctgcccct tggctgctct cggactccct accccaaccc cagggactgc aggttgtgcc
     3481 ctgtggctgc ctgcacccca ggaaaggagg gggctcaggc catccagcca ccacctacag
     3541 cccagtgggt acggtcgagg atatcgcggc cggccgcact cctcaggtgc aggctgccta
     3601 tcagaaggtg gtggctggtg tggccaatgc cctggctcac aaataccact gagatctttt
     3661 tccctctgcc aaaaattatg gggacatcat gaagcccctt gagcatctga cttctggcta
     3721 ataaaggaaa tttattttca ttgcaatagt gtgttggaat tttttgtgtc tctcactcgg
     3781 aaggacatat gggagggcaa atcatttaaa acatcagaat gagtatttgg tttagagttt
     3841 ggcaacatat gccatatgct ggctgccatg aacaaaggtg gctataaaga ggtcatcagt
     3901 atatgaaaca gccccctgct gtccattcct tattccatag aaaagccttg acttgaggtt
     3961 agattttttt tatattttgt tttgtgttat ttttttcttt aacatcccta aaattttcct
     4021 tacatgtttt actagccaga tttttcctcc tctcctgact actcccagtc atagctgtcc
     4081 ctcttctctt atgaagatcc ctcgacctgc agcccaagct tggcgtaatc atggtcatag
     4141 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc
     4201 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc
     4261 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc ggatccgcat ctcaattagt
     4321 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg
     4381 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
     4441 cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca
     4501 aaaagctaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     4561 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     4621 tgtatcttat catgtctgga tccgctgcat taatgaatcg gccaacgcgc ggggagaggc
     4681 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     4741 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     4801 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     4861 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     4921 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     4981 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     5041 gcctttctcc cttcgggaag cgtggcgctt tctcaatgct cacgctgtag gtatctcagt
     5101 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     5161 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     5221 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     5281 gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc
     5341 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     5401 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     5461 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     5521 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     5581 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     5641 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     5701 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     5761 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     5821 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     5881 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     5941 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     6001 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     6061 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     6121 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     6181 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     6241 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     6301 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     6361 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     6421 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     6481 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     6541 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     6601 ccgcgcacat ttccccgaaa agtgccacct g
//
