LOCUS       sequence-258284-        5208 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 5208)
  AUTHORS   Harding HP, Novoa I, Zhang Y, Zeng H, Wek R, Schapira M, Ron D
  TITLE     Regulated translation initiation controls stress-induced gene 
            expression in mammalian cells.
  JOURNAL   Mol Cell. 2000 Nov . 6(5):1099-108.
  PUBMED    11106749
REFERENCE   2  (bases 1 to 5208)
  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: "Mol Cell. 
            2000 Nov . 6(5):1099-108."
COMMENT     Sequence Label: ATF4 3: Mouse ATF4 (CHOP11/cATF), 5'UTR and AU...
FEATURES             Location/Qualifiers
     source          1..5208
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      267..276
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             478..2130
                     /codon_start=1
                     /gene="luc+"
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="enhanced luc+ version of the luciferase gene"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     primer_bind     complement(544..562)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    2171..2292
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2182..2201)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2236..2255
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2540..2557)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2711..3299)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2791..2810)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3470..4261,4262..4330))
                     /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     4093..4112
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4331..4435)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4462..4917
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4549..4568)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4759..4780
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    5048..5096
                     /note="synthetic polyadenylation signal"
     misc_feature    5110..5201
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     5150..5169
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ggtaccgagc tcttacgcgt gctagcccgg gctcgagatc tgacttctag agatccggca
       61 aaccccgccc agcgtcttgt cattggcgaa ttcgaacacg cagatgcagt cggggcggcg
      121 cggtccgagg tccacttcgc atattaaggt gacgcgtgtg gcctcgaaca ccgagcgacc
      181 ctgcataagc ttttctgctt gctgtctgcc ggtttaagtt gtgtgctcgg gtgtcccttt
      241 cctcttcccc tcccgcaggg cttgcggcca ccatggcgta ttagaggcag cagtgcctgc
      301 ggcagcgttg gcctttgcag cggcggcagc agcaccaggc tctgcagcgg caacccccac
      361 cggcctaggc catggcgctc ttcacgaaat ccagcagcag tgttgctgta acggacaaag
      421 ataccttcga gttaagcaca ttcctcgaat ccagcatagc cccacaacat gaccgtcatg
      481 gaagacgcca aaaacataaa gaaaggcccg gcgccattct atccgctgga agatggaacc
      541 gctggagagc aactgcataa ggctatgaag agatacgccc tggttcctgg aacaattgct
      601 tttacagatg cacatatcga ggtggacatc acttacgctg agtacttcga aatgtccgtt
      661 cggttggcag aagctatgaa acgatatggg ctgaatacaa atcacagaat cgtcgtatgc
      721 agtgaaaact ctcttcaatt ctttatgccg gtgttgggcg cgttatttat cggagttgca
      781 gttgcgcccg cgaacgacat ttataatgaa cgtgaattgc tcaacagtat gggcatttcg
      841 cagcctaccg tggtgttcgt ttccaaaaag gggttgcaaa aaattttgaa cgtgcaaaaa
      901 aagctcccaa tcatccaaaa aattattatc atggattcta aaacggatta ccagggattt
      961 cagtcgatgt acacgttcgt cacatctcat ctacctcccg gttttaatga atacgatttt
     1021 gtgccagagt ccttcgatag ggacaagaca attgcactga tcatgaactc ctctggatct
     1081 actggtctgc ctaaaggtgt cgctctgcct catagaactg cctgcgtgag attctcgcat
     1141 gccagagatc ctatttttgg caatcaaatc attccggata ctgcgatttt aagtgttgtt
     1201 ccattccatc acggttttgg aatgtttact acactcggat atttgatatg tggatttcga
     1261 gtcgtcttaa tgtatagatt tgaagaagag ctgtttctga ggagccttca ggattacaag
     1321 attcaaagtg cgctgctggt gccaacccta ttctccttct tcgccaaaag cactctgatt
     1381 gacaaatacg atttatctaa tttacacgaa attgcttctg gtggcgctcc cctctctaag
     1441 gaagtcgggg aagcggttgc caagaggttc catctgccag gtatcaggca aggatatggg
     1501 ctcactgaga ctacatcagc tattctgatt acacccgagg gggatgataa accgggcgcg
     1561 gtcggtaaag ttgttccatt ttttgaagcg aaggttgtgg atctggatac cgggaaaacg
     1621 ctgggcgtta atcaaagagg cgaactgtgt gtgagaggtc ctatgattat gtccggttat
     1681 gtaaacaatc cggaagcgac caacgccttg attgacaagg atggatggct acattctgga
     1741 gacatagctt actgggacga agacgaacac ttcttcatcg ttgaccgcct gaagtctctg
     1801 attaagtaca aaggctatca ggtggctccc gctgaattgg aatccatctt gctccaacac
     1861 cccaacatct tcgacgcagg tgtcgcaggt cttcccgacg atgacgccgg tgaacttccc
     1921 gccgccgttg ttgttttgga gcacggaaag acgatgacgg aaaaagagat cgtggattac
     1981 gtcgccagtc aagtaacaac cgcgaaaaag ttgcgcggag gagttgtgtt tgtggacgaa
     2041 gtaccgaaag gtcttaccgg aaaactcgac gcaagaaaaa tcagagagat cctcataaag
     2101 gccaagaagg gcggaaagat cgccgtgtaa ttctagagtc ggggcggccg gccgcttcga
     2161 gcagacatga taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa
     2221 aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc
     2281 aataaacaag ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggaggtg
     2341 tgggaggttt tttaaagcaa gtaaaacctc tacaaatgtg gtaaaatcga taaggatccg
     2401 tcgaccgatg cccttgagag ccttcaaccc agtcagctcc ttccggtggg cgcggggcat
     2461 gactatcgtc gccgcactta tgactgtctt ctttatcatg caactcgtag gacaggtgcc
     2521 ggcagcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     2581 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     2641 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     2701 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     2761 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     2821 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     2881 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     2941 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     3001 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     3061 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     3121 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     3181 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     3241 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     3301 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     3361 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     3421 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     3481 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     3541 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     3601 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     3661 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     3721 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     3781 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     3841 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     3901 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     3961 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     4021 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     4081 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     4141 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     4201 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     4261 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     4321 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     4381 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     4441 ttccccgaaa agtgccacct gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
     4501 tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt
     4561 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
     4621 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg
     4681 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
     4741 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct
     4801 cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg
     4861 agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgctt acaatttgcc
     4921 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
     4981 tacgccagcc caagctacca tgataagtaa gtaatattaa ggtacgggag gtacttggag
     5041 cggccgcaat aaaatatctt tattttcatt acatctgtgt gttggttttt tgtgtgaatc
     5101 gatagtacta acatacgctc tccatcaaaa caaaacgaaa caaaacaaac tagcaaaata
     5161 ggctgtcccc agtgcaagtg caggtgccag aacatttctc tatcgata
//
