LOCUS       sequence-14378-e        6188 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 6188)
  AUTHORS   Zito E, Melo EP, Yang Y, Wahlander A, Neubert TA, Ron D
  TITLE     Oxidative protein folding by an endoplasmic reticulum-localized 
            peroxiredoxin.
  JOURNAL   Mol Cell. 2010 Dec 10. 40(5):787-97.
  PUBMED    21145486
REFERENCE   2  (bases 1 to 6188)
  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. 
            2010 Dec 10. 40(5):787-97."
COMMENT     Sequence Label: hPDI1-18-508_C400S_C56S-pFLAG-CMV1
FEATURES             Location/Qualifiers
     source          1..6188
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     128..150
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     142..159
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     143..159
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        312..690
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        691..894
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     845..865
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             925..969
                     /codon_start=1
                     /product="leader sequence from mouse preprotrypsin"
                     /label=preprotrypsin leader
                     /translation="MSALLILALVGAAVA"
     CDS             970..993
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    2527..3151
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3173..3193)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3178..3508
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3359..3494
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3546..3565)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3547..3565)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3579..3595)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3579..3595)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3592..3614)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3603..3619
                     /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(3627..3633,3634..3651,3652..3657))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3672..3693
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3810..3827)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3981..4569)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4061..4080)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4740..5531,5532..5600))
                     /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     5363..5382
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5601..5705)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5732..6188
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5819..5838)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6029..6050
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 cccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc
       61 tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg gtaacgccca
      121 gggttttccc agtcacgacg ttgtaaaacg acggccagtg ccaagctgat ctaatcaata
      181 ttggccatta gccatattat tcattggtta tatagcataa atcaatattg gctattggcc
      241 attgcatacg ttgtatccat atcataatat gtacatttat attggctcat gtccaacatt
      301 accgccatgt tgacattgat tattgactag ttattaatag taatcaatta cggggtcatt
      361 agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg
      421 cgaccgccca gcgacccccg cccgttgacg tcaatagtga cgtatgttcc catagtaacg
      481 ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg
      541 gcagtacatc aagtgtatca tatgccaagt ccgcccccta ttgacgtcaa tgacggtaaa
      601 tggcccgcct agcattatgc ccagtacatg accttacggg agtttcctac ttggcagtac
      661 atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta caccaatggg
      721 cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg
      781 agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaataa ccccgccccg
      841 ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctcgttta
      901 gtgaaccgtc agaattaatt caccatgtct gcacttctga tcctagctct tgttggagct
      961 gcagttgctg actacaaaga cgatgacgac aagcttagcg acgcccccga ggaggaggac
     1021 cacgtcctgg tgctgaggaa aagcaacttc gcggaggcgc tggcggccca caagtacctg
     1081 ctggtggagt tctatgcccc ttggtgtggc cactccaagg ctctggcccc tgagtatgcc
     1141 aaagccgctg ggaagctgaa ggcagaaggt tccgagatca ggttggccaa ggtggacgcc
     1201 acggaggagt ctgacctggc ccagcagtac ggcgtgcgcg gctatcccac catcaagttc
     1261 ttcaggaatg gagacacggc ttcccccaag gaatatacag ctggcagaga ggctgatgac
     1321 atcgtgaact ggctgaagaa gcgcacgggc ccggctgcca ccaccctgcc tgacggcgca
     1381 gctgcagagt ccttggtgga gtccagcgag gtggctgtca tcggcttctt caaggacgtg
     1441 gagtcggact ctgccaagca gtttttgcag gcagcagagg ccatcgatga cataccattt
     1501 gggatcactt ccaacagtga cgtgttctcc aaataccagc tcgacaaaga tggggttgtc
     1561 ctctttaaga agtttgatga aggccggaac aactttgaag gggaggtcac caaggagaac
     1621 ctgctggact ttatcaaaca caaccagctg ccccttgtca tcgagttcac cgagcagaca
     1681 gccccgaaga tttttggagg tgaaatcaag actcacatcc tgctgttctt gcccaagagt
     1741 gtgtctgact atgacggcaa actgagcaac ttcaaaacag cagccgagag cttcaagggc
     1801 aagatcctgt tcatcttcat cgacagcgac cacaccgaca accagcgcat cctcgagttc
     1861 tttggcctga agaaggaaga gtgcccggcc gtgcgcctca tcaccctgga ggaggagatg
     1921 accaagtaca agcccgaatc ggaggagctg acggcagaga ggatcacaga gttctgccac
     1981 cgcttcctgg agggcaaaat caagccccac ctgatgagcc aggagctgcc ggaggactgg
     2041 gacaagcagc ctgtcaaggt gcttgttggg aagaactttg aagacgtggc ttttgatgag
     2101 aaaaaaaacg tctttgtgga gttctatgcc ccatggtgtg gccactccaa acagttggct
     2161 cccatttggg ataaactggg agagacgtac aaggaccatg agaacatcgt catcgccaag
     2221 atggactcga ctgccaacga ggtggaggcc gtcaaagtgc acagcttccc cacactcaag
     2281 ttctttcctg ccagtgccga caggacggtc atcgattaca acggggaacg cacgctggat
     2341 ggttttaaga aattcctgga gagcggtggc caggatgggg caggggatga tgacgatctc
     2401 gaggacctgg aagaagcaga ggagccagac atggaggaag acgatgatca gaaagctgtg
     2461 aaagatgaac tgtaaggatc cgagctcgag atctgcagct ggtaccagtc gactctagag
     2521 gatcccgggt ggcatccctg tgacccctcc ccagtgcctc tcctggcctt ggaagttgcc
     2581 actccagtgc ccaccagcct tgtcctaata aaattaagtt gcatcatttt gtctgactag
     2641 gtgtcctcta taatattatg gggtggaggg gggtggtatg gagcaagggg cccaagttgg
     2701 gaagacaacc tgtagggcct gcggggtcta ttcgggaacc aagctggagt gcagtggcac
     2761 aatcttggct cactgcaatc tccgcctcct gggttcaagc gattctcctg cctcagcctc
     2821 ccgagttgtt gggattccag gcatgcatga ccaggctcag ctaatttttg tttttttggt
     2881 agagacgggg tttcaccata ttggccaggc tggtctccaa ctcctaatct caggtgatct
     2941 acccaccttg gcctcccaaa ttgctgggat tacaggcgtg aaccactgct cccttccctg
     3001 tccttctgat tttaaaataa ctataccagc aggaggacgt ccagacacag cataggctac
     3061 ctgccatggc ccaaccggtg ggacatttga gttgcttgct tggcactgtc ctctcatgcg
     3121 ttgggtccac tcagtagatg cctgttgaat tgggtacgcg gccagcttct gtggaatgtg
     3181 tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     3241 catctcaatt agtcagcaac caggtgtgga aaagtcccca ggctccccag caggcagaag
     3301 tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat
     3361 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt
     3421 tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg
     3481 cttttttgga ggcctaggct tttgcaaaaa gctcctcgag gaactgaaaa accagaaagt
     3541 taattcccta tagtgagtcg tattaaattc gtaatcatgt catagctgtt tcctgtgtga
     3601 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
     3661 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
     3721 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     3781 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     3841 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     3901 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     3961 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     4021 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     4081 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     4141 gcctttctcc cttcgggaag cgtggcgctt tctcaatgct cacgctgtag gtatctcagt
     4201 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     4261 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     4321 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     4381 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     4441 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     4501 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     4561 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     4621 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     4681 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     4741 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     4801 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     4861 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     4921 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     4981 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     5041 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     5101 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     5161 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     5221 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     5281 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     5341 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     5401 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     5461 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     5521 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     5581 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     5641 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     5701 ccgcgcacat ttccccgaaa agtgccacct gacgcgccct gtagcggcgc attaagcgcg
     5761 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
     5821 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     5881 aatcggggca tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     5941 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     6001 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     6061 aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg
     6121 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaaacgtt
     6181 tacaattt
//
