LOCUS       sequence-128154-        5053 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 5053)
  AUTHORS   Chaudhry AZ, Vitullo AD, Gronostajski RM
  TITLE     Nuclear factor I (NFI) isoforms differentially activate simple 
            versus complex NFI-responsive promoters.
  JOURNAL   J Biol Chem. 1998 Jul 17. 273(29):18538-46.
  PUBMED    9660824
REFERENCE   2  (bases 1 to 5053)
  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: "J Biol 
            Chem. 1998 Jul 17. 273(29):18538-46."
COMMENT     Sequence Label: pCHNFI-B2
FEATURES             Location/Qualifiers
     source          1..5053
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        27..330
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        331..534
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     485..505
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     531..555
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          672..768
                     /label=SV40 intron
                     /note="modified SV40 intron with splice donor and acceptor 
                     sites"
     CDS             834..860
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    2192..2326
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2216..2235)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2270..2289
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2437..2453)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2437..2453)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2450..2472)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2461..2477
                     /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(2485..2491,2492..2509,2510..2515))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2530..2551
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2668..2685)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2839..3427)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2919..2938)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3598..4389,4390..4458))
                     /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     4221..4240
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4459..4563)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4631..4649
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4687..4709)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4809..4828
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5022..5044
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5036..5053
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5037..5053
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 gaattcgagc ttgcatgcct gcaggtcgtt acataactta cggtaaatgg cccgcctggc
       61 tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg
      121 ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg
      181 gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa
      241 tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac
      301 atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg
      361 cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg
      421 agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca
      481 ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctcgttta
      541 gtgaaccgtc agatcgcctg gagacgccat ccacgctgtt ttgacctcca tagaagacac
      601 cgggaccgat ccagcctccg gactctagag gatccggtac tcgaggaact gaaaaaccag
      661 aaagttaact ggtaagttta gtctttttgt cttttatttc aggtcccgga tccggtggtg
      721 gtgcaaatca aagaactgct cctcagtgga tgttgccttt acttctaggc ctgtacggaa
      781 gtgttacttc tgctctaaaa gctgcggaat tgtacccgcg gccgccagcc atgtatcctt
      841 acgatgtacc agactatgct gggcctcacg ggccgatgat gtattctccc atctgtctca
      901 ctcaggatga attccaccca tttattgagg cacttcttcc tcacgtccgt gcaattgcct
      961 atacttggtt caacctgcag gctcgaaaac gcaagtactt taaaaagcat gagaaacgaa
     1021 tgtcgaagga tgaagaaagg gcagtcaaag acgagctgct cagtgagaag cccgaaatca
     1081 agcagaagtg ggcatccagg ctcctggcca aactgcgcaa agatatccgc caggagtacc
     1141 gggaggactt tgtgcttacc gtgactggca agaagcaccc gtgctgtgtc ttatccaatc
     1201 cagaccagaa gggtaagatt aggaggatcg actgcctgcg acaggcagac aaagtctggc
     1261 gtctggatct agtcatggtg atcctgttca aaggcatccc tttggagagt acggatggag
     1321 agcgactcat gaagtccccg cactgcacaa acccagcact ttgtgttcag ccacaccaca
     1381 tcacagtatc agttaaggag cttgacttgt ttttggcata ctacgtgcag gagcaagatt
     1441 ctggacaatc aggaagtcca agccacagtg atcctgccaa gaatcctcca gggtacctcg
     1501 aggacagctt tgtaaaatcc ggagtcttca atgtatcaga gcttgtgaga gtatccagaa
     1561 cacccataac ccagggaact ggagtcaact tcccaatcgg agaaattccc agccaaccat
     1621 actatcatga catgaactct ggtgtgaacc tgcagaggtc gctgtcttct ccaccgagca
     1681 gcaaaagacc caaaactata tctatagatg aaaatatgga gccaagtcct acaggagact
     1741 tttacccctc tccaaattca ccagctgctg gaagtcgaac atggcatgaa cgagatcaag
     1801 atatgtcttc tccaactaca atgaagaagc ctgagaagcc actgtttagc tctacatctc
     1861 cacaggattc ttccccaaga ttgagcactt tcccccagca ccatcatccc ggaatacctg
     1921 gagtcgcgca cagtgtcatc tcaactcgaa ctccacctcc gccctcaccg ttgccatttc
     1981 cgacgcaagc tatccttcct ccggcacctt ccagctactt ctctcatcca acaatcagat
     2041 atcctcctca cctgaatcct caggatactc tgaagaacta cgtaccttct tatgacccat
     2101 ccagtcctca aacgagccag tcctggtacc tgggctagct tggttccttt tcaagtgtca
     2161 agatctatgc atagctagct agggccgcgg ggatccagac atgataagat acattgatga
     2221 gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga
     2281 tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca acaacaattg
     2341 cattcatttt atgtttcagg ttcaggggga ggtgtgggag gttttttcgg atcctctaga
     2401 gtcgacctgc aggcatgcaa gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     2461 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     2521 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     2581 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     2641 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     2701 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     2761 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     2821 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     2881 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     2941 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     3001 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     3061 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     3121 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     3181 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     3241 gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc
     3301 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     3361 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     3421 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     3481 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     3541 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     3601 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     3661 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     3721 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     3781 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     3841 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     3901 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     3961 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     4021 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     4081 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     4141 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     4201 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     4261 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     4321 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     4381 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     4441 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     4501 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     4561 gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt
     4621 aacctataaa aataggcgta tcacgaggcc ctttcgtctc gcgcgtttcg gtgatgacgg
     4681 tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt aagcggatgc
     4741 cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct
     4801 taactatgcg gcatcagagc agattgtact gagagtgcac catatgcggt gtgaaatacc
     4861 gcacagatgc gtaaggagaa aataccgcat caggcgccat tcgccattca ggctgcgcaa
     4921 ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg
     4981 atgtgctgca aggcgattaa gttgggtaac gccagggttt tcccagtcac gacgttgtaa
     5041 aacgacggcc agt
//
