LOCUS       sequence-10147-e        6046 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 6046)
  TITLE     ATF4 plasmids
REFERENCE   2  (bases 1 to 6046)
  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"
COMMENT     Sequence Label: ATF4.18: mATF4 (CHOP11/cATF)-GST
FEATURES             Location/Qualifiers
     source          1..6046
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(183..188,189..204,205..211)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    219..235
                     /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)."
     CDS             258..911
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     869..891
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             918..935
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     primer_bind     complement(2096..2118)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2263..2281)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2349..2453
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2454..2522,2523..3314)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2672..2691)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3485..4073
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3974..3993
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4227..4244
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        4317..4394
                     /gene="lacI (mutant)"
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the 
                     promoter boosts expression of the lacI gene about 10-fold."
     CDS             4395..5477
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPSTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(4414..4433)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     protein_bind    5490..5511
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5526..5531,5532..5549,5550..5556)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5564..5580
                     /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)."
     primer_bind     5588..5604
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5588..5604
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             join(5600..6046,1..81)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(5620..5637)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5620..5636)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5629..5651)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 agcttatcga ctgcacggtg caccaatgct tctggcgtca ggcagccatc ggaagctgtg
       61 gtatggctgt gcaggtcgta aatcactgca taattcgtgt cgctcaaggc gcactcccgt
      121 tctggataat gttttttgcg ccgacatcat aacggttctg gcaaatattc tgaaatgagc
      181 tgttgacaat taatcatcgg ctcgtataat gtgtggaatt gtgagcggat aacaatttca
      241 cacaggaaac agtattcatg tcccctatac taggttattg gaaaattaag ggccttgtgc
      301 aacccactcg acttcttttg gaatatcttg aagaaaaata tgaagagcat ttgtatgagc
      361 gcgatgaagg tgataaatgg cgaaacaaaa agtttgaatt gggtttggag tttcccaatc
      421 ttccttatta tattgatggt gatgttaaat taacacagtc tatggccatc atacgttata
      481 tagctgacaa gcacaacatg ttgggtggtt gtccaaaaga gcgtgcagag atttcaatgc
      541 ttgaaggagc ggttttggat attagatacg gtgtttcgag aattgcatat agtaaagact
      601 ttgaaactct caaagttgat tttcttagca agctacctga aatgctgaaa atgttcgaag
      661 atcgtttatg tcataaaaca tatttaaatg gtgatcatgt aacccatcct gacttcatgt
      721 tgtatgacgc tcttgatgtt gttttataca tggacccaat gtgcctggat gcgttcccaa
      781 aattagtttg ttttaaaaaa cgtattgaag ctatcccaca aattgataag tacttgaaat
      841 ccagcaagta tatagcatgg cctttgcagg gctggcaagc cacgtttggt ggtggcgacc
      901 atcctccaaa atcggatctg gttccgcgtg gatcccccgg gctgcaggat tcggcacgag
      961 ctcatgggtt ctccagcgac aaggcgggct cctcggaatg gccggctatg gatgatggct
     1021 tggccagtgc ctcagacacc ggcaaggagg atgccttttc cgggacagat tggatgttgg
     1081 agaaaatgga tctgaaagag tttgacttcg atgctctgtt tcgaatggat gacctggaaa
     1141 ccatgccaga tgagctcttg accacgttgg atgacacatg tgatcttttt gcccctctag
     1201 tccaagagac taataaggag ccccctcaga cagtgaaccc aattggccat ctcccagaaa
     1261 gtttaataaa agtcgaccag gttgccccct ttacattctt gcagcctttc ccctgttccc
     1321 caggggttct gtcttccact ccagagcatt cctttagttt agagctaggc agtgaagttg
     1381 atatctctga aggagacagg aagcctgact ctgctgctta cattactcta atccctccat
     1441 gtgtaaagga ggaagacact ccctctgaca atgacagtgg catctgtatg agcccggagt
     1501 cctacctggg ctctccccag catagcccct ccacctccag ggccccacca gacaatctgc
     1561 cttctccagg tggttcccgt gggtctcctc ggcccaaacc ttatgaccca cctggagtta
     1621 gtttgacagc taaagtgaag actgagaaat tggataagaa gctgaaaaag atggagcaaa
     1681 acaagacagc agccactagg taccgccaga agaagcgggc tgagcaggag gccctcactg
     1741 gcgagtgtaa ggagctagaa aaaaagaatg aggctctgaa agagaaggca gattctctgg
     1801 ccaaggagat ccagtatctg aaagacctga tagaagaggt ccgtaaggca agggggaaga
     1861 agagagttcc gtaatagggt agtcaggtgc tttgtgcttg tacatagtct tgtgttgctg
     1921 tgtttgctgt aataaattat tttgtagtga aagtaaaaaa aaaaaaaaaa aaaaaaaaaa
     1981 aaaaaaaaaa aaactcgagc cgaattctag actccatggg tcgactcgag ctcaagctta
     2041 attcacgtga ctgactgacg atctgcctcg cgcgtttcgg tgatgacggt gaaaacctct
     2101 gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac
     2161 aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggcgcagcc atgacccagt
     2221 cacgtagcga tagcggagtg tataattctt gaagacgaaa gggcctcgtg atacgcctat
     2281 ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg
     2341 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     2401 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
     2461 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     2521 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
     2581 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     2641 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtgttg
     2701 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     2761 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     2821 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
     2881 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     2941 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgcag
     3001 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
     3061 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     3121 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     3181 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     3241 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     3301 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
     3361 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     3421 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     3481 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     3541 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     3601 gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc
     3661 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     3721 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     3781 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     3841 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     3901 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     3961 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     4021 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     4081 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     4141 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg
     4201 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc
     4261 tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata aattccgaca
     4321 ccatcgaatg gtgcaaaacc tttcgcggta tggcatgata gcgcccggaa gagagtcaat
     4381 tcagggtggt gaatgtgaaa ccagtaacgt tatacgatgt cgcagagtat gccggtgtct
     4441 cttatcagac cgtttcccgc gtggtgaacc aggccagcca cgtttctgcg aaaacgcggg
     4501 aaaaagtgga agcggcgatg gcggagctga attacattcc caaccgcgtg gcacaacaac
     4561 tggcgggcaa acagtcgttg ctgattggcg ttgccacctc cagtctggcc ctgcacgcgc
     4621 cgtcgcaaat tgtcgcggcg attaaatctc gcgccgatca actgggtgcc agcgtggtgg
     4681 tgtcgatggt agaacgaagc ggcgtcgaag cctgtaaagc ggcggtgcac aatcttctcg
     4741 cgcaacgcgt cagtgggctg atcattaact atccgctgga tgaccaggat gccattgctg
     4801 tggaagctgc ctgcactaat gttccggcgt tatttcttga tgtctctgac cagacaccca
     4861 tcaacagtat tattttctcc catgaagacg gtacgcgact gggcgtggag catctggtcg
     4921 cattgggtca ccagcaaatc gcgctgttag cgggcccatt aagttctgtc tcggcgcgtc
     4981 tgcgtctggc tggctggcat aaatatctca ctcgcaatca aattcagccg atagcggaac
     5041 gggaaggcga ctggagtgcc atgtccggtt ttcaacaaac catgcaaatg ctgaatgagg
     5101 gcatcgttcc cactgcgatg ctggttgcca acgatcagat ggcgctgggc gcaatgcgcg
     5161 ccattaccga gtccgggctg cgcgttggtg cggatatctc ggtagtggga tacgacgata
     5221 ccgaagacag ctcatgttat atcccgccgt caaccaccat caaacaggat tttcgcctgc
     5281 tggggcaaac cagcgtggac cgcttgctgc aactctctca gggccaggcg gtgaagggca
     5341 atcagctgtt gcccgtctca ctggtgaaaa gaaaaaccac cctggcgccc aatacgcaaa
     5401 ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac
     5461 tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agctcactca ttaggcaccc
     5521 caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag cggataacaa
     5581 tttcacacag gaaacagcta tgaccatgat tacggattca ctggccgtcg ttttacaacg
     5641 tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt
     5701 cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag
     5761 cctgaatggc gaatggcgct ttgcctggtt tccggcacca gaagcggtgc cggaaagctg
     5821 gctggagtgc gatcttcctg aggccgatac tgtcgtcgtc ccctcaaact ggcagatgca
     5881 cggttacgat gcgcccatct acaccaacgt aacctatccc attacggtca atccgccgtt
     5941 tgttcccacg gagaatccga cgggttgtta ctcgctcaca tttaatgttg atgaaagctg
     6001 gctacaggaa ggccagacgc gaattatttt tgatggcgtt ggaatt
//
