LOCUS       sequence-381961-        5810 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Includes the promoter (1kb) of SMTS with 2 mutated KLF binding sites
            (position 449,732).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5810)
  TITLE     Leisegang-Brandes Plasmids
REFERENCE   2  (bases 1 to 5810)
  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"
COMMENT     Sequence Label: pGL3-pSMANTIS_1kb_1 KLF2mut 449,732
FEATURES             Location/Qualifiers
     source          1..5810
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      1074..1083
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1080..2732
                     /codon_start=1
                     /gene="luc+"
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="enhanced luc+ version of the luciferase gene"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     primer_bind     complement(1146..1164)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     polyA_signal    2773..2894
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2784..2803)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2838..2857
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3142..3159)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3313..3901)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3393..3412)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4072..4863,4864..4932))
                     /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     4695..4714
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4933..5037)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5064..5519
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5151..5170)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5361..5382
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    5650..5698
                     /note="synthetic polyadenylation signal"
     misc_feature    5712..5803
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     5752..5771
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ggtaccctat gtctgacaca tagtagagct taataaacat tttctattat tgctacggtt
       61 gttgttactg ttagcaacat tgaggctgag taaaatgaat tctttttgga tagaaagatg
      121 gtctcatggt caagtgtttt tacccaaatc tcaaggagat tctaaatagc agctaagaac
      181 caggggatac ttctctggcc aaaagtctac tgagacaagg tcttgggggc agaggctagg
      241 ccaggtagtt tgtggtgcac atagaatcac tgtacctacc cagcatgaag gtgttggccc
      301 ccaaggagct aactggaggg tagcatgctg gaagacacta ccagtgttat aggaggcaaa
      361 aagaaattat ttaggtagac atttagggta aagtgagtcc ccagcagaaa actttccttt
      421 taacaaaaag cagctcaaaa atagctccct ttctaatcat atatagttaa aaaaaatcac
      481 ttctcttcta acaacaagta acctgaaagt tcaggctgta aaatacagat aagacagctc
      541 aagcacagaa gaaggggtgg aagtctcctg ggtaattgcc aaacttcaca cttatacaat
      601 gggccccagt aaaacagtgg gtcttaataa gcacattcct ttccctttag gtgcactaag
      661 atagagaaac taaaagcaga ctcatggggc ggatgcctgc agctgcaaga agatgtatgg
      721 gaacagacac agaaactctt tctttcagat aagcaagaca aagaaacaca gactaagagt
      781 cagcctatgt ggtcagggaa tgggataaaa gctgataaaa aaaattctgc tctatataga
      841 tagcacactt ggtcgcagct aaaccatcag gccctaagag gataagacat cccctcctca
      901 tgaggcccct cctccctaga ccatttataa aaaccctgac atttttacta ctgcttagca
      961 acctgctcgg gacccctctc tgtgactgtg agctgttctg ttcttttgcc tagctagccc
     1021 gggctcgaga tctgcgatct aagtaagctt ggcattccgg tactgttggt aaagccacca
     1081 tggaagacgc caaaaacata aagaaaggcc cggcgccatt ctatccgctg gaagatggaa
     1141 ccgctggaga gcaactgcat aaggctatga agagatacgc cctggttcct ggaacaattg
     1201 cttttacaga tgcacatatc gaggtggaca tcacttacgc tgagtacttc gaaatgtccg
     1261 ttcggttggc agaagctatg aaacgatatg ggctgaatac aaatcacaga atcgtcgtat
     1321 gcagtgaaaa ctctcttcaa ttctttatgc cggtgttggg cgcgttattt atcggagttg
     1381 cagttgcgcc cgcgaacgac atttataatg aacgtgaatt gctcaacagt atgggcattt
     1441 cgcagcctac cgtggtgttc gtttccaaaa aggggttgca aaaaattttg aacgtgcaaa
     1501 aaaagctccc aatcatccaa aaaattatta tcatggattc taaaacggat taccagggat
     1561 ttcagtcgat gtacacgttc gtcacatctc atctacctcc cggttttaat gaatacgatt
     1621 ttgtgccaga gtccttcgat agggacaaga caattgcact gatcatgaac tcctctggat
     1681 ctactggtct gcctaaaggt gtcgctctgc ctcatagaac tgcctgcgtg agattctcgc
     1741 atgccagaga tcctattttt ggcaatcaaa tcattccgga tactgcgatt ttaagtgttg
     1801 ttccattcca tcacggtttt ggaatgttta ctacactcgg atatttgata tgtggatttc
     1861 gagtcgtctt aatgtataga tttgaagaag agctgtttct gaggagcctt caggattaca
     1921 agattcaaag tgcgctgctg gtgccaaccc tattctcctt cttcgccaaa agcactctga
     1981 ttgacaaata cgatttatct aatttacacg aaattgcttc tggtggcgct cccctctcta
     2041 aggaagtcgg ggaagcggtt gccaagaggt tccatctgcc aggtatcagg caaggatatg
     2101 ggctcactga gactacatca gctattctga ttacacccga gggggatgat aaaccgggcg
     2161 cggtcggtaa agttgttcca ttttttgaag cgaaggttgt ggatctggat accgggaaaa
     2221 cgctgggcgt taatcaaaga ggcgaactgt gtgtgagagg tcctatgatt atgtccggtt
     2281 atgtaaacaa tccggaagcg accaacgcct tgattgacaa ggatggatgg ctacattctg
     2341 gagacatagc ttactgggac gaagacgaac acttcttcat cgttgaccgc ctgaagtctc
     2401 tgattaagta caaaggctat caggtggctc ccgctgaatt ggaatccatc ttgctccaac
     2461 accccaacat cttcgacgca ggtgtcgcag gtcttcccga cgatgacgcc ggtgaacttc
     2521 ccgccgccgt tgttgttttg gagcacggaa agacgatgac ggaaaaagag atcgtggatt
     2581 acgtcgccag tcaagtaaca accgcgaaaa agttgcgcgg aggagttgtg tttgtggacg
     2641 aagtaccgaa aggtcttacc ggaaaactcg acgcaagaaa aatcagagag atcctcataa
     2701 aggccaagaa gggcggaaag atcgccgtgt aattctagag tcggggcggc cggccgcttc
     2761 gagcagacat gataagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa
     2821 aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct
     2881 gcaataaaca agttaacaac aacaattgca ttcattttat gtttcaggtt cagggggagg
     2941 tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg tggtaaaatc gataaggatc
     3001 cgtcgaccga tgcccttgag agccttcaac ccagtcagct ccttccggtg ggcgcggggc
     3061 atgactatcg tcgccgcact tatgactgtc ttctttatca tgcaactcgt aggacaggtg
     3121 ccggcagcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     3181 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     3241 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     3301 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     3361 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     3421 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     3481 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     3541 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     3601 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     3661 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     3721 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     3781 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     3841 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     3901 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     3961 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     4021 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     4081 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     4141 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     4201 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     4261 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     4321 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     4381 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     4441 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     4501 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     4561 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     4621 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     4681 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     4741 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     4801 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     4861 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     4921 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     4981 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     5041 atttccccga aaagtgccac ctgacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     5101 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
     5161 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     5221 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta
     5281 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     5341 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat
     5401 ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa
     5461 tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc ttacaatttg
     5521 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
     5581 attacgccag cccaagctac catgataagt aagtaatatt aaggtacggg aggtacttgg
     5641 agcggccgca ataaaatatc tttattttca ttacatctgt gtgttggttt tttgtgtgaa
     5701 tcgatagtac taacatacgc tctccatcaa aacaaaacga aacaaaacaa actagcaaaa
     5761 taggctgtcc ccagtgcaag tgcaggtgcc agaacatttc tctatcgata
//
