LOCUS       sequence-452307-        5176 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For subcloning of SV40 promoter or for assays using M13 phage.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5176)
  AUTHORS   DeMeis JD, Roberts JT, Delcher HA, Godang NL, Coley AB, Brown CL, 
            Shaw MH, Naaz S, Dahal A, Alqudah SY, Nguyen KN, Nguyen AD, Paudel 
            SS, Shell JE, Patil SS, Dang H, O'Neal WK, Knowles MR, Houserova D, 
            Gillespie MN, Borchert GM
  TITLE     Long G4-rich enhancers target promoters via a G4 DNA-based 
            mechanism.
  JOURNAL   Nucleic Acids Res. 2025 Jan 11;53(2):gkae1180. doi: 
            10.1093/nar/gkae1180.
  PUBMED    39658038
REFERENCE   2  (bases 1 to 5176)
  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: "Nucleic 
            Acids Res."; date: "2025-01-11"; volume: "53(2):gkae1180. doi"; 
            pages: " 10.1093/nar/gkae1180"
COMMENT     Sequence Label: TOPO-SV40
FEATURES             Location/Qualifiers
     source          1..5176
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    107..128
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(143..148,149..166,167..173)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    181..197
                     /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     186..208
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     205..221
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     205..221
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    350..398
                     /note="synthetic polyadenylation signal"
     CDS             complement(423..1022)
                     /codon_start=1
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     promoter        complement(1053..1410)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1067..1202
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(1121..1140)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     enhancer        1219..1410
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     1367..1387
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     primer_bind     complement(1609..1628)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1610..1628)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1635..1652)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1635..1651)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1644..1666)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      1792..2220
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1879..1898)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2089..2110
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             2564..3358
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2618..2637)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3228..3247
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     CDS             join(3376..3444,3445..4236)
                     /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     complement(3594..3613)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4407..4995
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4896..4915
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5149..5166
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
       61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
      121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
      181 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagcttg
      241 gtaccgagct cggatccact agtaacggcc gccagtgtgc tggaattcgc ccttgagctg
      301 actgggttga aggctctcaa gggcatcggt cgacagcatc tagttcgaac acacaaaaaa
      361 ccaacacaca gatgtaatga aaataaagat attttattgc ggccatgacc ggttctagat
      421 cattaggcgc caggttttcg tgtcatacac caggtcctag gtccctcggg cacctcgaca
      481 tcggcggtca cggtgaagcc caggcgctcg tagaatggaa ggtttcttgg tgcgcttgtt
      541 tcgagaaagg cgggcacacc ggcgcgttcg gcggcctcta cgccgggcaa cacgacggcg
      601 ctgcccaagc ccttgccctg gtggtcgggg ctcactccta cagtggccag aaaccaggca
      661 ggctccttgg gacgatgggg ggcaagcagg ccctccattt gctgctgggc ggccaggcgg
      721 ctgccgctca gctcggccat tcgggggccg atctcggcga acacggcgcc ggcctcaacg
      781 ctctcggggg tagtccacac ggccacggcc gcgccgtcgt cggctaccca caccttgccg
      841 atgtccagcc cgacgcgggt caggaacagc tcctgcaact ctgtcacacg ctcgatgtgc
      901 cggtcggggt ccacggtgtg ccgggtagcg gggtagtcgg cgaaagcggc ggccagagtg
      961 cggacggcgc ggggcacatc atcgcgagtg gccaggcgca cggtaggctt gtactcggtc
     1021 atggtggcgc tagtgtcaga agaatcgagc tttttgcaaa agcctaggcc tccaaaaaag
     1081 cctcctcact acttctggaa tagctcagag gcagaggcgg cctcggcctc tgcataaata
     1141 aaaaaaatta gtcagccatg gggcggagaa tgggcggaac tgggcggagt taggggcggg
     1201 atgggcggag ttaggggcgg gactatggtt gctgactaat tgagatgcat gctttgcata
     1261 cttctgcctg ctggggagcc tggggacttt ccacacctgg ttgctgacta attgagatgc
     1321 atgctttgca tacttctgcc tgctggggag cctggggact ttccacaccc taactgacac
     1381 acattccaca gctggttctt tccgcctcag aaggtagcta accaagttcc tctttcagag
     1441 gttatttcag gccatggtgc tgcgcagatc agcggaagag cgcccaatac gcaaacagat
     1501 ccttatcgat tttaccacat ttgtagaggt tttacttgca agggcgaatt ctgcagatat
     1561 ccatcacact ggcggccgct cgagcatgca tctagagggc ccaattcgcc ctatagtgag
     1621 tcgtattaca attcactggc cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt
     1681 acccaactta atcgccttgc agcacatccc cctttcgcca gctggcgtaa tagcgaagag
     1741 gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg gacgcgccct
     1801 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
     1861 ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
     1921 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
     1981 ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct
     2041 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
     2101 tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta taagggattt
     2161 tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
     2221 ttaacaaaat tcagggcgca agggctgcta aaggaagcgg aacacgtaga aagccagtcc
     2281 gcagaaacgg tgctgacccc ggatgaatgt cagctactgg gctatctgga caagggaaaa
     2341 cgcaagcgca aagagaaagc aggtagcttg cagtgggctt acatggcgat agctagactg
     2401 ggcggtttta tggacagcaa gcgaaccgga attgccagct ggggcgccct ctggtaaggt
     2461 tgggaagccc tgcaaagtaa actggatggc tttcttgccg ccaaggatct gatggcgcag
     2521 gggatcaaga tctgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg
     2581 attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca
     2641 acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt
     2701 tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg
     2761 gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga
     2821 agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatccca
     2881 ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct
     2941 tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac
     3001 tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc
     3061 gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt
     3121 gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt
     3181 catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg
     3241 tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat
     3301 cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgaat
     3361 tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
     3421 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
     3481 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
     3541 gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt
     3601 ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat
     3661 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg
     3721 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
     3781 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
     3841 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
     3901 cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa
     3961 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca
     4021 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
     4081 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
     4141 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
     4201 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat
     4261 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     4321 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     4381 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     4441 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     4501 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
     4561 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     4621 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     4681 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     4741 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     4801 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     4861 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     4921 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     4981 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     5041 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
     5101 gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg
     5161 agcgaggaag cggaag
//
