LOCUS       sequence-46926-e        5048 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 5048)
  AUTHORS   Bhakta MS, Segal DJ
  TITLE     The generation of zinc finger proteins by modular assembly.
  JOURNAL   Methods Mol Biol. 2010;649:3-30.
  PUBMED    20680825
REFERENCE   2  (bases 1 to 5048)
  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: "Methods Mol
            Biol."; date: "2010"; volume: "649"; pages: "3-30"
COMMENT     Sequence Label: pPGK-empty
FEATURES             Location/Qualifiers
     source          1..5048
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        15..515
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     434..452
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     promoter        664..682
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     664..681
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     695..714
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     CDS             761..787
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     regulatory      1296..1305
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    1309..1443
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1346..1365)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1400..1419
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        1512..1869
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        1512..1703
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(1535..1555)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      1720..1855
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(1889..1906)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1889..1905)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1898..1920)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2118..2573
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2205..2224)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2415..2436
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2590..2609)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2744..2766
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2819..2837)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2905..3009
                     /gene="bla"
                     /label=AmpR promoter
     CDS             3195..3573
                     /label=ISS
                     /note="immunostimulatory sequence from the AmpR gene; 
                     contains unmethylated CpG dinucleotides in the context of 
                     5'-AACGTT-3' (Sato et al., 1996)"
     primer_bind     complement(3229..3248)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4042..4630
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4531..4550
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4784..4801
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        join(4954..4959,4960..4977,4978..4984)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4992..5008
                     /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     4997..5019
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5016..5032
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5016..5032
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 tcgagctctt ggggttgcgc cttttccaag gcagccctgg gtttgcgcag ggacgcggct
       61 gctctgggcg tggttccggg aaacgcagcg gcgccgaccc tgggtctcgc acattcttca
      121 cgtccgttcg cagcgtcacc cggatcttcg ccgctaccct tgtgggcccc ccggcgacgc
      181 ttcctgctcc gcccctaagt cgggaaggtt ccttgcggtt cgcggcgtgc cggacgtgac
      241 aaacggaagc cgcacgtctc actagtaccc tcgcagacgg acagcgccag ggagcaatgg
      301 cagcgcgccg accgcgatgg gctgtggcca atagcggctg ctcagcgggg cgcgccgaga
      361 gcagcggccg ggaaggggcg gtgcgggagg cggggtgtgg ggcggtagtg tgggccctgt
      421 tcctgcccgc gcggtgttcc gcattctgca agcctccgga gcgcacgtcg gcagtcggct
      481 ccctcgttga ccgaatcacc gacctctctc cccagggccg cggccgggaa cggtgcattg
      541 gaacgcggat tccccgtgcc aagagtgacg taagtaccgc ctatagagtc tatggcccac
      601 ccccttggct tcgttagaac gcggctacaa ttaatacata accttatgta tcatacacat
      661 acgatttagg tgacactata gaataacatc cactttgcct ttctctccac aggtgtccac
      721 tcccaggtcc aactgcacct cggttctatc gattgacatg tacccatacg atgtcccaga
      781 ctacgcgaat tcttgaaatg aaggtaatgg aattttttat gaaagtttat ggatatagag
      841 gtaaacattt gggtggatca aggaaaccgg acggagcaat ttatactgtc ggatctccta
      901 ttgattacgg tgtgatcgtg gatactaaag cttatagcgg aggttataat ctgccaattg
      961 gccaagcaga tgaaatgcaa gattatgtcg aagaaaatca aacacgaaac aaacatatca
     1021 accctaatga atggtggaaa gtctatccat cttctgtaac ggaatttaag tttttatttg
     1081 tgagtggtca ctttaaagga aactacaaag ctcagcttac acgattaaat catatcacta
     1141 attgtaatgg agctgttctt agtgtagaag agcttttaat tggtggagaa atgattaaag
     1201 ccggcacatt aaccttagag gaagtgagac ggaaatttaa taacggcgag ataaactttt
     1261 aattaactaa tctagagtcg acctgcagaa gcttggccgc catggcccaa cttgtttatt
     1321 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     1381 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg
     1441 atcgggaatt aattcggcgc agcaccatgg cctgaaataa cctctgaaag aggaacttgg
     1501 ttaggtacct tctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga
     1561 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     1621 accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     1681 aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc
     1741 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
     1801 gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc
     1861 atttgcaaaa agctgttaac agcttggcac tggccgtcgt tttacaacgt cgtgactggg
     1921 aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca tccccccttc gccagctggc
     1981 gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgtagc ctgaatggcg
     2041 aatggcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatac
     2101 gtcaaagcaa ccatagtacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     2161 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
     2221 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
     2281 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg atttgggtga
     2341 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     2401 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggg
     2461 ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct
     2521 gatttaacaa aaatttaacg cgaattttaa caaaatatta acgtttacaa ttttatggtg
     2581 cactctcagt acaatctgct ctgatgccgc atagttaagc caactccgct atcgctacgt
     2641 gactgggtca tggctgcgcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct
     2701 tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt
     2761 cagaggtttt caccgtcatc accgaaacgc gcgaggcagt attcttgaag acgaaagggc
     2821 ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca
     2881 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     2941 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     3001 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
     3061 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
     3121 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat cctttgagag
     3181 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
     3241 ggtattatcc cgtgatgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     3301 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
     3361 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     3421 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     3481 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     3541 caccacgatg ccagcagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
     3601 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
     3661 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     3721 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     3781 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     3841 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     3901 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     3961 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     4021 agaaaagatc aaaggatctt cttcagatcc tttttttctg cgcgtaatct gctgcttgca
     4081 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     4141 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc ttctagtgta
     4201 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     4261 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     4321 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     4381 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agcattgaga
     4441 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     4501 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     4561 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     4621 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     4681 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
     4741 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
     4801 ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta
     4861 atccagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa
     4921 tgtgagttac ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat
     4981 gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta
     5041 cgaattaa
//
