LOCUS       sequence-455934-        6554 bp    DNA     circular SYN 15-APR-2025
DEFINITION  For subcloning of human CEP72 promoter or for assays using M13 
            phage.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6554)
  TITLE     Borchert Lab TOPO plasmids
REFERENCE   2  (bases 1 to 6554)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: TOPO-CEP72
FEATURES             Location/Qualifiers
     source          1..6554
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    79..100
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(115..120,121..138,139..145)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    153..169
                     /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     158..180
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     177..193
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     177..193
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2959..2978)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2960..2978)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2985..3002)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2985..3001)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2994..3016)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3142..3570
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3229..3248)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3439..3460
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             3914..4708
                     /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(3968..3987)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4578..4597
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     CDS             join(4726..4794,4795..5586)
                     /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(4944..4963)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5757..6345
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6246..6265
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6499..6516
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg
       61 cagtgagcgc aacgcaatta atgtgagtta gctcactcat taggcacccc aggctttaca
      121 ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc ggataacaat ttcacacagg
      181 aaacagctat gaccatgatt acgccaagct tggtaccgag ctcggatcca ctagtaacgg
      241 ccgccagtgt gctggaattc gcccttctgc atacacccca tgtacaccct ctgtctgtgt
      301 ataccctcgt ctactaccac catacgtacc ccccaccact accctgcctt gtctatcacc
      361 ccccaggctg tgacatacca gggatgttgc caacgagctg gtctaatctg gggtgagggt
      421 ttctgggtgc ccctagacac caggagaaag gcggggaggg gacacccctg ggggtctgga
      481 ttcactccac caggcgctgg gccacagtgg tacggtgaca cagtagatgt cttgagcagg
      541 attaaagtct tcccaataaa caggctgaac tctgtcttcc ttgaaagctc tgtagaagga
      601 cctcacaagg ataaacacag cgagaccctc cagggcgcgg tcagccacga gccacagtgc
      661 tgggcctgtg acatctctgg aggttctggg gtggagaagg accggccgtg gggagtgact
      721 ggctggggag aagctcagct cccgcccacc tctgggaggc ccggggctgg acgggaggtg
      781 cagcagacaa ggctccgggc gggtgctggg gtgtctggag gggtccccgc gggggggtcg
      841 ggttgggtgg ggagtctagg tgagtcgggg gtccccgggg gcgctgggtt cagggtggga
      901 gcggagtgtg gatggggcag ggtccccatg cccaccacca gccgcgattc caccgcgcct
      961 actcggggtt tccccgggag gacaccgcgt gtgtcaaggg ccggggccta gaaggcggcc
     1021 gcgcccaggg aagccttgac agggagctat ctggggcggg ggctggggtc gcgtggccag
     1081 tacctgggag aaccgccatt caggagaacc ccccgccagg ctcttggttc cgcccacgca
     1141 gtcccgcccc ctcccccagc cacgcccctc gccccgccgc gcaggccccg ccccgtcccg
     1201 ccgggctctc ggttctgccg acgcaggtcc cgcccctctc gtagcctcgc cccgcccgcc
     1261 gcgcaggcgc cgtccgaggg ctccgtttgt aacatggcgc gggctggccc tcggctggtg
     1321 ctgagcgagg aggcggttcg ggcgaagagc ggcttagggc ctcaccgcga cctgggtgcg
     1381 ccggagggcg ggcgggggtg caagcgtgag gtggcggggg ggtgggtgcc gagcttcccg
     1441 gggcggcggc ggaccgtggg caggcgggac ccgtctacgc aggcgccgcg ggcgtccgga
     1501 acggtcgcgg gcgagcgggg cgcgcgtgtc caggtgagcg acccaccccc cgtgcccagg
     1561 tgcggcccct gcccgcgttc gggtcccggc gtcccggccc ctgcctgtct atcgggccac
     1621 tggttccgtg actgggggtt acatccgctg agaagaggga gccggccttg ctgtccagga
     1681 gcgcagcccg ccctcccgct ggggatcacc tccctggttc tcctgggccc actctgtggt
     1741 gattgttgcg ctcggccttc ttgaggatgt tggtggccgg tttccttctt tttggcttat
     1801 ttttgtctgg ttttggcgtc aaggtgaagc cgagctcata aactgctgag aagtgttgcc
     1861 ttctcttata tttagaagag gttctgtaga gtgtgtgctc attgtttgaa tgtttcctgg
     1921 aattttccag tgaaaccata tggacatgtc aatttctgtt tttgaagctt ttaaatgaca
     1981 aattctgtat ctctggttgt tagagaaccg ttcatattag ttcaccgcga gtcttggctg
     2041 tgcggggttt tgtggcattg gcttatttga gctaacttgt ggggtgtgtg tgttgagttg
     2101 tttctggtgt tctgttgtta gcctgttaat gtctgtgtgt ggtctgcagg tgttactgga
     2161 ccaacaggtt cagtcaggtg ctgtgggggc cacaggccaa tacactgaga cagtggggtt
     2221 tgcagcggag aaagagttta aggctggcag gggcccagca gggaggtgga aggaggcctc
     2281 aatccatctc cccaaggaga tctgagcggt tttttttttt ttagacggag tctcactctg
     2341 tcgtccaggt tggagtgcag tggcacaatc tcagctcact gcaacctcca cctcccaggt
     2401 tcaagtgatt cactcagcct cctgagtagc tgggattaca agcacgcgcc accatgcccg
     2461 gctaattttt tatattttta gtagagacgg ggtttcacca tgttggccag gctggtcttg
     2521 aactcctgat ctcaggtgat ccacctgcct tggcctccca aagtgctggg attacaggca
     2581 tgagccacct acacccagcc tgagctgggg gttttagagg aacaggactg tgaggagctg
     2641 gaagattggt gttgtcagtt ggtggaggta agggccatgg aattatcagg gtgtggaaac
     2701 tgcattcttt ggtgaatccg cttgcggggc cctttagacc agctggcatc agtccccagg
     2761 tcctgaaaga atatctcaaa tggaaaactt aacggtttat aatgtccagg ttattatctg
     2821 tagagtggtt aaggggaact gtaacctcgt gacaaaatcc aagtgatcct gaggcagtag
     2881 gcaccacaga agggcgaatt ctgcagatat ccatcacact ggcggccgct cgagcatgca
     2941 tctagagggc ccaattcgcc ctatagtgag tcgtattaca attcactggc cgtcgtttta
     3001 caacgtcgtg actgggaaaa ccctggcgtt acccaactta atcgccttgc agcacatccc
     3061 cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc ccaacagttg
     3121 cgcagcctga atggcgaatg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
     3181 tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt
     3241 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
     3301 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg
     3361 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
     3421 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct
     3481 cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg
     3541 agctgattta acaaaaattt aacgcgaatt ttaacaaaat tcagggcgca agggctgcta
     3601 aaggaagcgg aacacgtaga aagccagtcc gcagaaacgg tgctgacccc ggatgaatgt
     3661 cagctactgg gctatctgga caagggaaaa cgcaagcgca aagagaaagc aggtagcttg
     3721 cagtgggctt acatggcgat agctagactg ggcggtttta tggacagcaa gcgaaccgga
     3781 attgccagct ggggcgccct ctggtaaggt tgggaagccc tgcaaagtaa actggatggc
     3841 tttcttgccg ccaaggatct gatggcgcag gggatcaaga tctgatcaag agacaggatg
     3901 aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt
     3961 ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt
     4021 gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc
     4081 cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc
     4141 ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga
     4201 agtgccgggg caggatctcc tgtcatccca ccttgctcct gccgagaaag tatccatcat
     4261 ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca
     4321 agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga
     4381 tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc
     4441 gcgcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat
     4501 catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga
     4561 ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg
     4621 ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt
     4681 ctatcgcctt cttgacgagt tcttctgaat tgaaaaagga agagtatgag tattcaacat
     4741 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
     4801 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
     4861 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
     4921 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg
     4981 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
     5041 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
     5101 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
     5161 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
     5221 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca
     5281 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
     5341 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
     5401 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
     5461 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
     5521 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
     5581 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
     5641 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     5701 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     5761 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     5821 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     5881 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     5941 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     6001 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     6061 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     6121 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     6181 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     6241 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     6301 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     6361 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     6421 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
     6481 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc
     6541 aaaccgcctc tccc
//
