LOCUS       sequence-55255-e        6626 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 6626)
  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 6626)
  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: rGH-PAL
FEATURES             Location/Qualifiers
     source          1..6626
                     /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      333..342
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(339..341,342..344,345..1058)
                     /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(339..341,342..344,345..1055)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(384..405)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(645..664)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     992..1013
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    1071..1638
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    1088..1640
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1232..1249)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1459..1478
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             1652..3151
                     /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(3650..3669)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3715..3770
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3769..3788)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4129..4145)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4129..4145)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4142..4164)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4153..4169
                     /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(4177..4183,4184..4201,4202..4207))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4222..4243
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4302..4497
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4348..4483
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    4503..4637
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4540..4559)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4594..4613
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4705..4722)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4876..5464)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4956..4975)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5635..6426,6427..6495))
                     /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     6258..6277
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6496..6600)
                     /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 cgagtgaggt catgacctca
      241 ccagcttcga attctatata atggaagctt gaccggagcc gttagttgcc cgtcggacgc
      301 gatttgcgag gcgctttatg cgcgcaccgg tcgccaccat ggtgagcaag ggcgaggagc
      361 tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt
      421 tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagttca
      481 tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ctgacctacg
      541 gcgtgcagtg cttcagccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg
      601 ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac ggcaactaca
      661 agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg
      721 gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac aactacaaca
      781 gccacaacgt ctatatcatg gccgacaagc agaagaacgg catcaaggtg aacttcaaga
      841 tccgccacaa catcgaggac ggcagcgtgc agctcgccga ccactaccag cagaacaccc
      901 ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagcacc cagtccgccc
      961 tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg
     1021 ccgggatcac tctcggcatg gacgagctgt acaagtaaag cggccaattc tccctccccc
     1081 ccccctaacg ttactggccg aagccgcttg gaataaggcc ggtgtgcgtt tgtctatatg
     1141 ttattttcca ccatattgcc gtcttttggc aatgtgaggg cccggaaacc tggccctgtc
     1201 ttcttgacga gcattcctag gggtctttcc cctctcgcca aaggaatgca aggtctgttg
     1261 aatgtcgtga aggaagcagt tcctctggaa gcttcttgaa gacaaacaac gtctgtagcg
     1321 accctttgca ggcagcggaa ccccccacct ggcgacaggt gcctctgcgg ccaaaagcca
     1381 cgtgtataag atacacctgc aaaggcggca caaccccagt gccacgttgt gagttggata
     1441 gttgtggaaa gagtcaaatg gctctcctca agcgtattca acaaggggct gaaggatgcc
     1501 cagaaggtac cccattgtat gggatctgat ctggggcctc ggtgcacatg ctttacgtgt
     1561 gtttagtcga ggttaaaaaa acgtctaggc cccccgaacc acggggacgt ggttttcctt
     1621 tgaaaaacac gatgataata tggggtcgag gctgctgctg ctgctgctgc tgggcctgag
     1681 gctacagctc tccctgggca tcatcctagt tgaggaggag aacccggact tctggaaccg
     1741 cgaggcagcc gaggccctgg gtgccgccaa gaagctgcag cctgcacaga cagccgccaa
     1801 gaacctcatc atcttcctgg gcgatgggat gggggtgtct acggtgacag ctgccaggat
     1861 cctaaaaggg cagaagaagg acaaactggg gcctgagata cccctggcca tggaccgctt
     1921 cccatatgtg gctctgtcca agacatacaa tgtagacaaa catgtgccag acagtggagc
     1981 cacagccacg gcctacctgt gcggggtcaa gggcaacttc cagaccattg gcttgagtgc
     2041 agccgcccgc tttaaccagt gcaacacgac acgcggcaac gaggtcatct ccgtgatgaa
     2101 tcgggccaag aaagcaggga agtcagtggg agtggtaacc accacacgag tgcagcacgc
     2161 ctcgccagcc ggcacctacg cccacacggt gaaccgcaac tggtactcgg acgccgacgt
     2221 gcctgcctcg gcccgccagg aggggtgcca ggacatcgct acgcagctca tctccaacat
     2281 ggacattgat gtgatcctag gtggaggccg aaagtacatg tttcgcatgg gaaccccaga
     2341 ccctgagtac ccagatgact acagccaagg tgggaccagg ctggacggga agaatctggt
     2401 gcaggaatgg ctcggcgaac gccagggtgc ccggtacgtg tggaaccgca ctgagctcat
     2461 gcaggcttcc ctggacccgt ctgtgaccca tctcatgggt ctctttgagc ctggagacat
     2521 gaaatacgag atccaccgag actccacact ggacccctcc ctgatggaga tgacagaggc
     2581 tgccctgcgc ctgctgagca gacacccccg cggcttcttc ctcttcgtgg agggtggtcg
     2641 catcgaccat ggtcatcatg aaagcagggc ttaccgggca ctgactgaga cgatcatgtt
     2701 cgacgacgcc attgagaggg cgggccagct caccagcgag gaggacacgc tgagcctcgt
     2761 cactgccgac cactcccacg tcttctcctt cggaggctac cccctgcgag ggagctcctt
     2821 catcgggctg gccgctggca aggcccggga caggaaggcc tacacggtcc tcctatacgg
     2881 aaacggtcca ggctatgtgc tcaaggacgg cgcccggccg gatgttaccg agagcgagag
     2941 cgggagcccc gagtatcggc agcagtcagc agtgcccctg gacgaagaga cccacgcagg
     3001 cgaggacgtg gcggtgttcg cgcgcggccc gcaggcgcac ctggttcacg gcgtgcagga
     3061 gcagaccttc atagcgcacg tcatggcctt cgccgcctgc ctggagccct acaccgcctg
     3121 cgacctggcg ccccccgccg gcaccaccga cgccgcgcac ccggggcggt ccgtggtccc
     3181 cgcgttgctt cctctgctgg ccgggaccct gctgctgctg gagacggcca ctgctccctg
     3241 agtgtcccgt ccctggggct cctgcttccc catcccggag ttctcctgct ccccacctcc
     3301 tgtcgtcctg cctggcctcc agcccgagtc gtcatccccg gagtccctat acagaggtcc
     3361 tgccatggaa ccttcccctc cccgtgcgct ctggggactg agcccatgac accaaacctg
     3421 ccccttggct gctctcggac tccctacccc aaccccaggg actgcaggtt gtgccctgtg
     3481 gctgcctgca ccccaggaaa ggagggggct caggccatcc agccaccacc tacagcccag
     3541 tgggtacggt cgaggatatc gcggccggcc gcactcctca ggtgcaggct gcctatcaga
     3601 aggtggtggc tggtgtggcc aatgccctgg ctcacaaata ccactgagat ctttttccct
     3661 ctgccaaaaa ttatggggac atcatgaagc cccttgagca tctgacttct ggctaataaa
     3721 ggaaatttat tttcattgca atagtgtgtt ggaatttttt gtgtctctca ctcggaagga
     3781 catatgggag ggcaaatcat ttaaaacatc agaatgagta tttggtttag agtttggcaa
     3841 catatgccat atgctggctg ccatgaacaa aggtggctat aaagaggtca tcagtatatg
     3901 aaacagcccc ctgctgtcca ttccttattc catagaaaag ccttgacttg aggttagatt
     3961 ttttttatat tttgttttgt gttatttttt tctttaacat ccctaaaatt ttccttacat
     4021 gttttactag ccagattttt cctcctctcc tgactactcc cagtcatagc tgtccctctt
     4081 ctcttatgaa gatccctcga cctgcagccc aagcttggcg taatcatggt catagctgtt
     4141 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     4201 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     4261 gcccgctttc cagtcgggaa acctgtcgtg ccagcggatc cgcatctcaa ttagtcagca
     4321 accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat
     4381 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc
     4441 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag
     4501 ctaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     4561 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     4621 cttatcatgt ctggatccgc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     4681 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     4741 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     4801 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     4861 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     4921 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     4981 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     5041 tctcccttcg ggaagcgtgg cgctttctca atgctcacgc tgtaggtatc tcagttcggt
     5101 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     5161 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     5221 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     5281 cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct
     5341 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     5401 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     5461 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     5521 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     5581 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     5641 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     5701 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     5761 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     5821 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     5881 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     5941 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     6001 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     6061 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     6121 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     6181 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     6241 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     6301 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     6361 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     6421 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     6481 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     6541 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     6601 cacatttccc cgaaaagtgc cacctg
//
