LOCUS       sequence-246519-        7099 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 7099)
  TITLE     ATF4 plasmids
REFERENCE   2  (bases 1 to 7099)
  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 1: Mouse ATF4 (CHOP11/cATF)-WT
FEATURES             Location/Qualifiers
     source          1..7099
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(2266..2284)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2267..2284)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2304..2321)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2310..2534
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2580..3008
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2667..2686)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2877..2898
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3017..3037)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3022..3351
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3202..3337
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3264..3283
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    4772..4893
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4809..4828)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4863..4882
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4942..4958)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4942..4958)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4955..4977)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4966..4982
                     /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(4990..4996,4997..5014,5015..5020))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5035..5056
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5173..5190)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5344..5932)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5424..5443)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6103..6894,6895..6963))
                     /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     6726..6745
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6964..7068)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gcttggtacc
      901 gagctcggat ccactagtaa cggccgccag tgtgctggaa ttcggcacga gacgaaatcc
      961 agcagcagtg ttgctgtaac ggacaaagat accttcgagt taagcacatt cctggaatcc
     1021 agcaaagccc cacaacatga ccgagatgag cttcctgaac agcgaagtgt tggcggggga
     1081 cttgatgtcc cccttcgacc agtcgggttt gggggctgaa gaaagcctag gtctcttaga
     1141 tgactatctg gaggtggcca agcacttgaa acctcatggg ttctccagcg acaaggcggg
     1201 ctcctcggaa tggccggcta tggatgatgg cttggccagt gcctcagaca ccggcaagga
     1261 ggatgccttt tccgggacag attggatgtt ggagaaaatg gatctgaaag agtttgactt
     1321 cgatgctctg tttcgaatgg atgacctgga aaccatgcca gatgagctct tgaccacgtt
     1381 ggatgacaca tgtgatcttt ttgcccctct agtccaagag actaataagg agccccctca
     1441 gacagtgaac ccaattggcc atctcccaga aagtttaata aaagtcgacc aggttgcccc
     1501 ctttacattc ttgcagcctt tcccctgttc cccaggggtt ctgtcttcca ctccagagca
     1561 ttcctttagt ttagagctag gcagtgaagt tgatatctct gaaggagaca ggaagcctga
     1621 ctctgctgct tacattactc taatccctcc atgtgtaaag gaggaagaca ctccctctga
     1681 caatgacagt ggcatctgta tgagcccgga gtcctacctg ggctctcccc agcatagccc
     1741 ctccacctcc agggccccac cagacaatct gccttctcca ggtggttccc gtgggtctcc
     1801 tcggcccaaa ccttatgacc cacctggagt tagtttgaca gctaaagtga agactgagaa
     1861 attggataag aagctgaaaa agatggagca aaacaagaca gcagccacta ggtaccgcca
     1921 gaagaagcgg gctgagcagg aggccctcac tggcgagtgt aaggagctag aaaaaaagaa
     1981 tgaggctctg aaagagaagg cagattctct ggccaaggag atccagtatc tgaaagacct
     2041 gatagaagag gtccgtaagg caagggggaa gaagagagtt ccgtaatagg gtagtcaggt
     2101 gctttgtgct tgtacatagt cttgtgttgc tgtgtttgct gtaataaatt attttgtagt
     2161 gaaagtaaaa aaaaaaaaaa aaaaaaacct cgtgccgctc gtgccgaatt ctgcagatat
     2221 ccatcacact ggcggccgct cgagcatgca tctagagggc cctattctat agtgtcacct
     2281 aaatgctaga gctcgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt
     2341 ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta
     2401 ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg
     2461 ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggatgc
     2521 ggtgggctct atggcttctg aggcggaaag aaccagctgg ggctctaggg ggtatcccca
     2581 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     2641 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     2701 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     2761 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
     2821 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     2881 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
     2941 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     3001 cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca
     3061 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc
     3121 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata
     3181 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     3241 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc tgcctctgag
     3301 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccg
     3361 ggctcgacta gagatccctc gacctcgaga tccattgtgc tctaaagcct aagatgagct
     3421 ttccatgtaa atttgtagcc agcttccttc tgattttcaa tgtttcttcc aaaggtgcag
     3481 tctccaaaga gattacgaat gccttggaaa cctggggtgc cttgggtcag gacatcaact
     3541 tggacattcc tagttttcaa atgagtgatg atattgacga tataaaatgg gaaaaaactt
     3601 cagacaagaa aaagattgca caattcagaa aagagaaaga gactttcaag gaaaaagata
     3661 catataagct atttaaaaat ggaactctga aaattaagca tctgaagacc gatgatcagg
     3721 atatctacaa ggtatcaata tatgatacaa aaggaaaaaa tgtgttggaa aaaatatttg
     3781 atttgaagat tcaagagagg gtctcaaaac caaagatctc ctggacttgt atcaacacaa
     3841 ccctgacctg tgaggtaatg aatggaactg accccgaatt aaacctgtat caagatggga
     3901 aacatctaaa actttctcag agggtcatca cacacaagtg gaccaccagc ctgagtgcaa
     3961 aattcaagtg cacagcaggg aacaaagtca gcaaggaatc cagtgtcgag cctgtcagct
     4021 gtccagagaa aggtctggac atctatctca tcattggcat atgtggagga ggcagcctct
     4081 tgatggtctt tgtggcactg ctcgttttct atatcaccaa aaggaaaaaa cagaggagtc
     4141 ggagaaatga tgaggagctg gagacaagag cccacagagt agctactgaa gaaaggggcc
     4201 ggaagcccca acaaattcca gcttcaaccc ctcagaatcc agcaacttcc caacatcctc
     4261 ctccaccacc tggtcatcgt tcccaggcac ctagtcatcg tcccccgcct cctggacacc
     4321 gtgttcagca ccagcctcag aagaggcctc ctgctccgtc gggcacacaa gttcaccagc
     4381 agaaaggccc gcccctcccc agacctcgag ttcagccaaa acctccccat ggggcagcag
     4441 aaaactcatt gtccccttcc tctaattaaa aaagatagaa actgtctttt tcaataaaaa
     4501 gcactgtgga tttctgccct cctgatgtgc atatccgtac ttccatgagg tgttttctgt
     4561 gtgcagaaca ttgtcacctc ctgaggctgt gggccacagc cacctctgca tcttcgaaat
     4621 gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta
     4681 tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg
     4741 ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta
     4801 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     4861 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag
     4921 ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
     4981 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
     5041 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     5101 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     5161 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     5221 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     5281 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     5341 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     5401 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     5461 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     5521 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     5581 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     5641 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     5701 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     5761 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     5821 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     5881 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     5941 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     6001 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     6061 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     6121 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     6181 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     6241 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     6301 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     6361 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     6421 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     6481 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     6541 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     6601 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     6661 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     6721 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     6781 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     6841 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     6901 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     6961 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     7021 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     7081 aaaagtgcca cctgacgtc
//
