LOCUS       sequence-14391-e        5499 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 5499)
  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 5499)
  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-A4
FEATURES             Location/Qualifiers
     source          1..5499
                     /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    2638..2772
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2662..2681)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2716..2735
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2883..2899)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2883..2899)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2896..2918)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2907..2923
                     /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(2931..2937,2938..2955,2956..2961))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2976..2997
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3114..3131)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3285..3873)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3365..3384)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4044..4835,4836..4904))
                     /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     4667..4686
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4905..5009)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5077..5095
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5133..5155)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5255..5274
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5468..5490
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5482..5499
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5483..5499
                     /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 ttctccgctc tgtctcaccc
      901 aggatgagtt tcatcctttc attgaagcac ttctgcccca tgtccgcgcc ttcgcctaca
      961 catggttcaa cctgcaggcc cgaaagcgga aatacttcaa aaaacatgag aagcgcatgt
     1021 cgaaagaaga ggagagggcc gtgaaggatg aactgctaag cgagaagccc gaggtcaagc
     1081 agaagtgggc ttcccgactt ctggccaagt tacggaaaga tatccgaccc gagtaccgag
     1141 aggattttgt tcttacagtt acagggaaaa aacctccatg ctgtgttctt tccaaccctg
     1201 atcagaaagg caagatgcgg agaattgact gcctccgcca ggcagataaa gtatggaggt
     1261 tggacctcgt catggtgatc ttgttcaaag gtattccgct ggaaagtact gatggcgagc
     1321 gccttgtaaa gagtccacag tgctctaatc cagggctctg tgtccagccc catcacatag
     1381 gggtttctgt aaaggaactc gatttatatt tggcatactt tgtacatgca gcagattcaa
     1441 gtcaatctga aagtcccagc cagccaagtg aagctgacat taaggaccag ccagaaaatg
     1501 gacatttggg cttccaggac agctttgtca catcaggtgt tttcagtgtg actgagctag
     1561 taagagtgtc acaaacacca atagctgcag gaaccggccc caatttttct ctctctgatt
     1621 tggaaagttc ttcatactac agcatgagtc caggagcaat gaggaggtct ctgcccagca
     1681 catcctctac cagctctaca aagcgcctca agtctgtgga ggatgaaatg gacagtcctg
     1741 gtgaagaacc attttacaca ggccaagggc gctccccagg gagtggcagc cagtccagtg
     1801 gatggcatga agtagagcca ggcttgccat ctccaagcac actgaagaag tctgagaagt
     1861 ctggtttcag cagcccctcc ccttcgcaga cctcctccct gggaacagca ttcacacagc
     1921 atcaccgacc tgtcattaca ggacccagag caagtccaca tgcgacgcca tcgactctcc
     1981 actttccaac gtcacccatc atccagcagc ctgggcctta cttctcacac ccagccatcc
     2041 gttaccaccc tcaggagacg ctgaaagagt ttgtccaact tgtctgtcct gatgctggtc
     2101 agcaagctgg acaggtgggg ttcctcaatc ccaatggaag cagtcaaggc aaggtgcaca
     2161 acccattcct ccccacccca atgttgccgc cgccgccacc accaccgatg gccaggcctg
     2221 tgcctctgcc catgccagac accaagcctc caaccacatc aacagaagga ggtgcagcct
     2281 cccccacctc accgatcctg gtacctggga taaaagttgc agcatcccac catcctccag
     2341 acagaccacc tgaccccttc tcaactctgt aacatggacg caacctcaac ccggcgcagt
     2401 tacaacttca ctgtcagtgg aaggggagcg aaaccaactc gaatcaactt atacatggaa
     2461 acagcaaaca tcgtggtcaa acagcaaagg ccataacttt ttgggattct ttttttttct
     2521 ttttttcttt ttttcttttt ttttttcttt tttaatactt tagggactgt cgtaatttct
     2581 catggtgctg gaaatggttg ggctgtgtga catttgaagt ctagctaggg ccgcggggat
     2641 ccagacatga taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa
     2701 aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc
     2761 aataaacaag ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggaggtg
     2821 tgggaggttt tttcggatcc tctagagtcg acctgcaggc atgcaagctt ggcgtaatca
     2881 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
     2941 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
     3001 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
     3061 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     3121 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     3181 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     3241 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     3301 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     3361 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     3421 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     3481 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     3541 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     3601 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     3661 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     3721 agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     3781 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     3841 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     3901 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     3961 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     4021 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     4081 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     4141 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     4201 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     4261 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     4321 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     4381 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     4441 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     4501 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     4561 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     4621 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     4681 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     4741 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     4801 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     4861 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     4921 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     4981 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
     5041 taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt
     5101 cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagacg
     5161 gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg
     5221 ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat cagagcagat tgtactgaga
     5281 gtgcaccata tgcggtgtga aataccgcac agatgcgtaa ggagaaaata ccgcatcagg
     5341 cgccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg
     5401 ctattacgcc agctggcgaa agggggatgt gctgcaaggc gattaagttg ggtaacgcca
     5461 gggttttccc agtcacgacg ttgtaaaacg acggccagt
//
