LOCUS       sequence-14389-e        4995 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 4995)
  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 4995)
  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-X2
FEATURES             Location/Qualifiers
     source          1..4995
                     /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    2134..2268
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2158..2177)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2212..2231
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2379..2395)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2379..2395)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2392..2414)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2403..2419
                     /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(2427..2433,2434..2451,2452..2457))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2472..2493
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2610..2627)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2781..3369)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2861..2880)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3540..4331,4332..4400))
                     /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     4163..4182
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4401..4505)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4573..4591
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4629..4651)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4751..4770
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4964..4986
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4978..4995
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4979..4995
                     /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 ggccgatgta tagcccgtac tgcctcaccc
      901 aggatgagtt ccacccgttt atcgaggcgc tgctgcctca cgtccgagcc ttctcctaca
      961 cctggttcaa cctgcaggcg cggaagcgca agtacttcaa gaagcacgag aagcggatgt
     1021 caaaggacga ggagcgcgca gtgaaggacg agctgctggg cgagaagcct gagatcaagc
     1081 agaagtgggc atcccggctg ttggccaagc tgcgcaaaga catccggccc gagttccgcg
     1141 aggactttgt gctaaccatc acgggcaaga agcccccctg ctgcgtgctt tccaaccccg
     1201 accagaaggg caagatccgg cggattgact gcctgcgcca ggctgacaag gtgtggcggc
     1261 tggacctggt catggtgatt ttgtttaaag ggatcccttt ggaaagtact gatggggagc
     1321 ggctctacaa gtcgccccag tgctcgaacc ccggcctgtg tgtccagcca catcacattg
     1381 gagtcacaat caaagaactg gacctttatc tggcttactt tgtccacact ccggaatccg
     1441 gacaatcaga tagttcaaac cagcaaggag atgcggacat caaaccactg cccaacgggc
     1501 acttaagttt ccaggactgc tttgtgacgt ctggggtctg gaatgtgaca gagctggtga
     1561 gagtatcaca gactccagtt gcgactgcat cagggcccaa cttctcactg gcggacctgg
     1621 agagccccag ctactacaac ataaatcaag tgaccctggg aaggcggtcc atcacctccc
     1681 ctccttccac cagcagcacc aagcgcccca agtccatcga cgacagtgag atggagagtc
     1741 cagtagatga tgtgttctat cctgggacag gccgctctcc ggccgctggc agcagccagt
     1801 ctagcggatg gcccaatgac gtggatgcag ggagcccccg ggccacggca tcagcgctgc
     1861 acttcccttc cacgtccatc atccagcagt cgagcccgta tttcacacac ccaaccatcc
     1921 gctaccacca ccaccatggg caggactcgc tgaaggagtt tgtgcagttt gtgtgctctg
     1981 acggctcggg tcaggccacc ggacagcatt cacaacgaca ggcgcctcct ctgccaaccg
     2041 gtttgtcagc atcggatccc gggacggcaa ctttctgaac atcccacagg agtctcagtc
     2101 ctagatctat gcatagctag ctagggccgc ggggatccag acatgataag atacattgat
     2161 gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt
     2221 gatgctattg ctttatttgt aaccattata agctgcaata aacaagttaa caacaacaat
     2281 tgcattcatt ttatgtttca ggttcagggg gaggtgtggg aggttttttc ggatcctcta
     2341 gagtcgacct gcaggcatgc aagcttggcg taatcatggt catagctgtt tcctgtgtga
     2401 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
     2461 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
     2521 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     2581 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     2641 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     2701 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     2761 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     2821 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     2881 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     2941 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     3001 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     3061 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     3121 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     3181 gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc
     3241 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     3301 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     3361 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     3421 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     3481 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     3541 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     3601 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     3661 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     3721 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     3781 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     3841 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     3901 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     3961 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     4021 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     4081 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     4141 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     4201 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     4261 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     4321 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     4381 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     4441 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     4501 ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca
     4561 ttaacctata aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac
     4621 ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct gtaagcggat
     4681 gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg
     4741 cttaactatg cggcatcaga gcagattgta ctgagagtgc accatatgcg gtgtgaaata
     4801 ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc attcgccatt caggctgcgc
     4861 aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg
     4921 ggatgtgctg caaggcgatt aagttgggta acgccagggt tttcccagtc acgacgttgt
     4981 aaaacgacgg ccagt
//
