LOCUS       sequence-238582-        6237 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression vector for Tre..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6237)
  AUTHORS   Bessen JL, Afeyan LK, Dancik V, Koblan LW, Thompson DB, Leichner C, 
            Clemons PA, Liu DR
  TITLE     High-resolution specificity profiling and off-target prediction for 
            site-specific DNA recombinases.
  JOURNAL   Nat Commun. 2019 Apr 26;10(1):1937. doi: 10.1038/s41467-019-09987-0.
  PUBMED    31028261
REFERENCE   2  (bases 1 to 6237)
  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: "Nat 
            Commun."; date: "2019-04-26"; volume: "10(1):1937. doi"; pages: " 
            10.1038/s41467-019-09987-0"
COMMENT     Sequence Label: pCMV-Tre
FEATURES             Location/Qualifiers
     source          1..6237
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        105..484
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        485..688
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     639..659
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     733..752
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        733..751
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     rep_origin      1974..2402
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2061..2080)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2271..2292
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2411..2431)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2416..2745
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2596..2731
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2658..2677
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2812..3606
                     /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(2866..2885)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3476..3495
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3780..3901
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3817..3836)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3871..3890
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3950..3966)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3950..3966)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3963..3985)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3974..3990
                     /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(3998..4004,4005..4022,4023..4028))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4043..4064
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4181..4198)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4352..4940)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4432..4451)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5111..5902,5903..5971))
                     /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     5734..5753
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5972..6076)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6151..6170)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc ttagggttag
       61 gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt gattattgac
      121 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
      181 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      241 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      301 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      361 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      421 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      481 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      541 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      601 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      661 acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca ctgcttactg
      721 gcttatcgaa attaatacga ctcactatag ggagacccaa gcttggtacc gagctcggat
      781 ccagccacca tgagcaattt actgaccctg caccatagtt tgcctgcatt accggccgat
      841 gcaacgagtg atgaggttcg caagaacctg atggacgtgt tcagggatcg cccggcgttt
      901 tctgagcata cctgggaaat gcttctgtcc gtttgccggt cgtgggcggc atggtgcaag
      961 ttgaataacc ggaaatggtt tcccgcagaa cctgaagatg ttcgcgatta tcttctacat
     1021 cttcaggcgc gcggtctggc agtaaaaact atccagcaac atttgtgcag gctaaacatg
     1081 cttcatcgtc ggtccgggct gccacgacca agtgacagca atgctgtttc actggttatg
     1141 cggcggatcc gaaaagaaaa cgttgatgcc ggtgaacgta ccaaacaggc tctagcgttc
     1201 gaacgcactg atttcgacca ggttcgttca ctcatggaaa atagcgatcg ctgccaggat
     1261 atacgtaatc tggcatttct gggggttgct tataacaccc tgttacgtat agccgaaatt
     1321 gccaggatca gggttaaaga tatctcacgt actgacggtg ggagaatgtt aatccatatt
     1381 ggcagaacga aaacgctggt tagcaccgca ggtgtagaga aggcacttag cctgggggta
     1441 actaaactgg tcgagcgatg gatttccgtc tctggtgtag ctgatgatcc gaataactac
     1501 ctgttttgcc gggtcagacg ttatggtgtt gccgcgccat ctgccaccag ccagctatca
     1561 acttacgccc tgcagcgcat ttttgaagca actcatcgat tgatttacgg cgctaaggat
     1621 gactctggtc agagatacct ggcctggtct ggacacagtg cccgtgtcgg agccgcgcga
     1681 gatatggccc gcgctggagt ttcaataccg gagatcatgc aagctggtgg ctggaccacc
     1741 gtaaatagcg tcatgaacta tatccgtaac ctggatagtg aaacaggggc aatggtgcgc
     1801 ctgctggaag atggcgacta agcggccgct cgagcatgca tctagaaata attcttactg
     1861 tcatgccaag taagatgctt ttctgtgctg caatagcagg catgctgggg atgcggtggg
     1921 ctctatggct tctgaggcgg aaagaaccag ctggggctct agggggtatc cccacgcgcc
     1981 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     2041 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     2101 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
     2161 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc
     2221 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     2281 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat
     2341 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     2401 ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc
     2461 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc
     2521 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg
     2581 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     2641 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc
     2701 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct
     2761 tgtatatcca ttttcggatc tgatcaagag acaggatgag gatcgtttcg catgattgaa
     2821 caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac
     2881 tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg
     2941 cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag
     3001 gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt
     3061 gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg
     3121 tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg
     3181 catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga
     3241 gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag
     3301 gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat
     3361 ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt
     3421 tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg
     3481 gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt
     3541 tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc
     3601 ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac
     3661 gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg
     3721 acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca
     3781 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     3841 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     3901 atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt
     3961 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     4021 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     4081 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     4141 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     4201 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     4261 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     4321 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     4381 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     4441 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     4501 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     4561 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     4621 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     4681 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     4741 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     4801 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     4861 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     4921 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     4981 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     5041 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     5101 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     5161 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     5221 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     5281 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     5341 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     5401 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     5461 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     5521 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     5581 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     5641 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     5701 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     5761 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     5821 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     5881 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     5941 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     6001 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     6061 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac ggatcgggag
     6121 atctcccgat cccctatggt gcactctcag tacaatctgc tctgatgccg catagttaag
     6181 ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag tagtgcgcga gcaaaat
//
