LOCUS       sequence-10017-e        6844 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 6844)
  TITLE     PERK constructs
REFERENCE   2  (bases 1 to 6844)
  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: PERK 9: PERK.pBSKS
FEATURES             Location/Qualifiers
     source          1..6844
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(33..461)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(143..164)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     355..374
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     588..610
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     602..619
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     603..619
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     626..645
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        626..644
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     677..693
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     677..693
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(4613..4629)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4614..4630)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     promoter        complement(4659..4677)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4698..4714)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4698..4714)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4711..4733)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4722..4738
                     /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(4746..4752,4753..4770,4771..4776))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4791..4812
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4929..4946)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5100..5688)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5180..5199)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5859..6650,6651..6719))
                     /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     6482..6501
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6720..6824)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cacctaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag
       61 ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac
      121 cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga
      181 ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc
      241 accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg
      301 gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa
      361 gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac
      421 caccacaccc gccgcgctta atgcgccgct acagggcgcg tcccattcgc cattcaggct
      481 gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa
      541 agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg
      601 ttgtaaaacg acggccagtg aattgtaata cgactcacta tagggcgaat tggagctcca
      661 ccgcggtggc ggccgctcta gaactagtgg atcgatcccc cgggctgcag gaattcggca
      721 cgagcgatgt ctgcacaagg ctgtcactca ggtggcagtg gctgacacgt ggccgggcag
      781 ctctgctgct gcggcgcgaa gtcgagaggc ggcggggtcc gtggcgcgcg ctcgcattgc
      841 tccgaggctc cgagcggcga tacgggcggg cgccgacggc agggtctcca tgcccgcgcg
      901 tggggcgggc cgctgatgga gcgcgccacc cggcccgggc cgcgcgcgct gctgctgctt
      961 ctgttcctgc tgctgggctg cgcggcgggg atctctgcgg tcgcgcccgc ccgcagtttg
     1021 cttgctcccg cgtcggagac agtgtttggc ttaggggcag cggccgcccc gacttcggcc
     1081 gcgcgggtgc ctgcggtggc aacggccgaa gtgaccgtgg aggacgccga ggcattgccg
     1141 gctgccgctg gcgaaccgga gtcacgcgcg acggagcccg atgacgacgt ggaactgcgg
     1201 cctcgcggca ggtccttggt aatcatcagc actttagatg gacgaatcgc tgcactggat
     1261 gccgagaatg atgggaaaaa gcagtgggat ttggacgtgg ggtctggttc cttggtttca
     1321 tctagcctca gcaagccaga ggtgtttggg aacaagatga tcatcccctc cctggatgga
     1381 gacctcttcc agtgggaccg ggaccgagag agcatggagg ccgtcccctt cacggtggag
     1441 tccctgctcg aatcttccta caagtttgga gatgatgttg ttctggttgg agggaaatct
     1501 ctgattacat acggactcag tgcttacagt ggaaagctga ggtatatctg ttctgccttg
     1561 ggatgtcgcc gatgggatag tgatgaaatg gaagaagagg aagacatctt gcttctgcag
     1621 cgtacgcaga agactgtgcg agctgtcggg cctcgaagcg gcagtgagaa gtggaatttc
     1681 agtgttggcc actttgaact tcggtatatt ccagacatgg aaactagagc cggattcatt
     1741 gaaagcacct ttaaaccggg tggaaacaaa gaagactcta aaattatttc agatgtggaa
     1801 gaacaagaag ccaccatgct ggacacagtg ataaaagttt ccgttgctga ttggaaggtc
     1861 atggcgttta gtaggaaggg aggccgcctg gaatgggagt accagttttg tactcccatc
     1921 gcgtccgcct ggctggtgag ggatggcaag gtcatcccca tcagcctgtt tgatgataca
     1981 agttacacag ccagcgaaga agccttggga gacgaagaag acattgtaga ggctgctcgg
     2041 ggagccacag agaacagcgt gtacttaggg atgtacagag gccagctgta cctgcagtcg
     2101 tccgtcaggg tctcagaaaa gttccctaca agcccaaagg ccttggagtc tgtaaatggc
     2161 gaaaatgcaa ttattcctct gccgacgatc aaatggaagc ccttaatcca ttctccttct
     2221 aggactcctg tcttggttgg gtctgatgaa tttgacaaat gtctaagtaa tgataagtat
     2281 tcccacgaag aatacagtaa tggtgcactt tcaatcctcc agtatccata cgataacggt
     2341 tactatctgc catactacaa gagagaaagg aataagcgga gcacgcagat cacagtcagg
     2401 ttcctggaca gcccccacta cagcaagaac atccgcaaga aggaccctat cctcctgctg
     2461 cactggtgga aggagatatt cgggacgatc ctgctttgca tcgtagccac gaccttcatc
     2521 gtgcgcaggc ttttccatcc tcagccccac aggcagcgga aggagtctga aactcagtgc
     2581 cagactgaaa gtaaatacga ctccgtgagt gccgatgtca gtgacaacag ctggaatgac
     2641 atgaagtact caggatacgt atcccgatat ctaacagatt ttgagccaat tcagtgcatg
     2701 ggtcgtggtg gctttggcgt tgtctttgaa gctaaaaaca aagtagatga ctgcaattac
     2761 gctatcaaga ggatccggct ccccaacagg gagttggcac gggagaaggt aatgcgggaa
     2821 gttaaagcct tggctaagct ggaacaccca ggcattgtga ggtatttcaa cgcctggctg
     2881 gaaaccccac cagagaagtg gcaagaagag atggatgaga tctggctcaa agacgaaagc
     2941 acagactggc cgctcagctc ccctagcccg atggatgccc catctgttaa gatccgaagg
     3001 atggatccnt tctctacaaa agagcagatc gaagtcatag ctccttctcc tgaaagaagt
     3061 cggtctttct cggtgggcat ttcctgtggc cagacaagct catcggagag ccagttctct
     3121 cccctggagt tctcagggac agactgcgga gacaacagtg actcagcgga cgcagcctac
     3181 aacctccagg acagttgcct gacggactgc gaggacgtgg aagatggcac cgtggacggc
     3241 aatgacgagg gacactcctt tgaactttgt ccgtccgaag cttctcccta tacccggtct
     3301 agggaaggaa cgtcctcctc catagtgttt gaggactctg gctgcggcaa cgcgtccagt
     3361 aaggaggagc ccagagggaa ccggctgcat gatggcaacc attatgttaa taagctaact
     3421 gatctcaagt gctccagcag caggtcttct tcagaagcca ccaccttgtc tacctcccct
     3481 accaggccaa ccactctaag cttggatttc accaagaaca ctgtgggcca gctccagccc
     3541 agctccccca aggtgtatct gtacattcag atgcagctgt gcaggaagga gaacctcaaa
     3601 gactggatga accggcgctg cagcttggag gaccgggagc acggcgtgtg cctgcacatc
     3661 ttcctgcaga tcgcagaggc agtggagttc ctgcacagca agggactcat gcacagggac
     3721 ctcaagcctt ccaacatatt cttcacaatg gatgatgtgg tcaaggttgg ggactttgga
     3781 ctggtgactg ctatggacca agatgaagaa gagcagactg ttctgactcc aatgccagcc
     3841 tatgctacgc acacgggaca agtagggacc aagctataca tgagcccaga gcagattcat
     3901 ggaaacaact actcccataa agtggacatc ttctctttag gcttgattct gtttgaactc
     3961 ctctacccat tcagcaccca gatggaacga gtccggattt taactgatgt cagaaatctc
     4021 aagtttcctc tactgttcac tcagaaatat ccccaagagc atatgatggt tcaagacatg
     4081 ctctctccat cccccacgga gcggcctgaa gccacagaca tcattgaaaa tgccatattt
     4141 gagaacttgg agtttcccgg gaaaacggtt ctgagacagc ggtcccgctc catgagttca
     4201 tctggaacaa aacattccag acagcccagc tgctcgtaca gcccactgcc tggcaactag
     4261 ccctcagctg ccctcgaagg tggcagagca ggcaccctga ggaacatggc tctccacagc
     4321 ggtggactca gattttatgc tttgatcagt tggactcggg accaattttt ctaagtcaga
     4381 ctggatcacg ggcctaaccc agtttgatct taactgaact tcaaggaaag ggctgtgtaa
     4441 aggacacatg aacttgttgc ttgtcggtgt cccaagacta gctggtcagc ttagaacctt
     4501 cacttttcac caggcggtag aagagatcct caaatggtct gaactggaaa tgtctttaaa
     4561 gcacaaaagt gtaaaagacc ctctcacatg ggaactacat gttctagctt atcgataccg
     4621 tcgacctcga gggggggccc ggtacccagc ttttgttccc tttagtgagg gttaatttcg
     4681 agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
     4741 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
     4801 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     4861 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     4921 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     4981 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     5041 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     5101 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     5161 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     5221 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     5281 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     5341 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     5401 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     5461 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     5521 aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc
     5581 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     5641 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     5701 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     5761 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     5821 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     5881 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     5941 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     6001 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     6061 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     6121 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     6181 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     6241 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     6301 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     6361 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     6421 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     6481 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     6541 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     6601 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     6661 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     6721 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     6781 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     6841 gtgc
//
