LOCUS       sequence-6886-ex        7501 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 7501)
  TITLE     Shivdasani lab plasmids
REFERENCE   2  (bases 1 to 7501)
  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: p079 pPNT6
FEATURES             Location/Qualifiers
     source          1..7501
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(52..85)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        108..604
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     CDS             618..1421
                     /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="MGSAIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQ
                     GRPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQD
                     LLSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE
                     EHQGLAPAELFARLKARMPDGDDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQ
                     DIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(681..700)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1291..1310
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    1437..1898
                     /label=PGK poly(A) signal
                     /note="mouse phosphoglycerate kinase 1 polyadenylation 
                     signal"
     protein_bind    complement(1916..1949)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        2024..2520
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     5'UTR           2562..2601
                     /label=HSV TK 5'-UTR
                     /note="5' untranslated region from the herpes simplex virus
                     thymidine kinase gene"
     CDS             2619..3749
                     /codon_start=1
                     /product="herpes simplex virus thymidine kinase"
                     /label=HSV TK
                     /note="confers sensitivity to nucleoside analogs such as 
                     acyclovir or ganciclovir"
                     /translation="MASYPCHQHASAFDQAARSRGHNNRRTALRPRRQQKATEVRLEQK
                     MPTLLRVYIDGPHGMGKTTTTQLLVALGSRDDIVYVPEPMTYWRVLGASETIANIYTTQ
                     HRLDQGEISAGDAAVVMTSAQITMGMPYAVTDAVLAPHIGGEAGSSHAPPPALTLIFDR
                     HPIAALLCYPAARYLMGSMTPQAVLAFVALIPPTLPGTNIVLGALPEDRHIDRLAKRQR
                     PGERLDLAMLAAIRRVYGLLANTVRYLQGGGSWREDWGQLSGAAVPPQGAEPQSNAGPR
                     PHIGDTLFTLFRAPELLAPNGDLYNVFAWALDVLAKRLRPMHVFILDYDQSPAGCRDAL
                     LQLTSGMVQTHVTTPGSIPTICDLARTFAREMGEAN"
     primer_bind     complement(3747..3766)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3791..3839
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     complement(4818..4835)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4818..4834)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4827..4849)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(5043..5062)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5162..5184
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5222..5240)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5308..5412
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5413..5481,5482..6273)
                     /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(5631..5650)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6444..7032
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6933..6952
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7186..7203
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7320..7341
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7356..7361,7362..7379,7380..7386)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7394..7410
                     /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     7399..7421
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7418..7434
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7418..7434
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        7455..7473
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 gcggccgcac gcgtcgacag cgtttaaacg aggtaccagt ggcgcgccgg cataacttcg
       61 tatagcatac attatacgaa gttataggcc tgcaggtcaa ttctaccggg taggggaggc
      121 gcttttccca aggcagtctg gagcatgcgc tttagcagcc ccgctggcac ttggcgctac
      181 acaagtggcc tctggcctcg cacacattcc acatccaccg gtagcgccaa ccggctccgt
      241 tctttggtgg ccccttcgcg ccaccttcta ctcctcccct agtcaggaag ttcccccccg
      301 ccccgcagct cgcgtcgtgc aggacgtgac aaatggaagt agcacgtctc actagtctcg
      361 tgcagatgga cagcaccgct gagcaatgga agcgggtagg cctttggggc agcggccaat
      421 agcagctttg ctccttcgct ttctgggctc agaggctggg aaggggtggg tccgggggcg
      481 ggctcagggg cgggctcagg ggcggggcgg gcgcgaaggt cctcccgagg cccggcattc
      541 tcgcacgctt caaaagcgca cgtctgccgc gctgttctcc tcttcctcat ctccgggcct
      601 ttcgacctgc agccaatatg ggatcggcca ttgaacaaga tggattgcac gcaggttctc
      661 cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct
      721 ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg
      781 acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg tggctggcca
      841 cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc
      901 tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga
      961 aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc
     1021 cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc
     1081 ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg
     1141 ccaggctcaa ggcgcgcatg cccgacggcg atgatctcgt cgtgacccat ggcgatgcct
     1201 gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc
     1261 tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc
     1321 ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc
     1381 agcgcatcgc cttctatcgc cttcttgacg agttcttctg aggggatcga tccgtcctgt
     1441 aagtctgcag aaattgatga tctattaaac aataaagatg tccactaaaa tggaagtttt
     1501 tcctgtcata ctttgttaag aagggtgaga acagagtacc tacattttga atggaaggat
     1561 tggagctacg ggggtggggg tggggtggga ttagataaat gcctgctctt tactgaaggc
     1621 tctttactat tgctttatga taatgtttca tagttggata tcataattta aacaagcaaa
     1681 accaaattaa gggccagctc attcctccca ctcatgatct atagatctat agatctctcg
     1741 tgggatcatt gtttttctct tgattcccac tttgtggttc taagtactgt ggtttccaaa
     1801 tgtgtcagtt tcatagcctg aagaacgaga tcagcagcct ctgttccaca tacacttcat
     1861 tctcagtatt gttttgccaa gttctaattc catcagaagc tgactctaga ggcctataac
     1921 ttcgtatagc atacattata cgaagttatg cgcaggatcc ccggttaacg ccttaattaa
     1981 ctcctcgagc atgctagcgt cgatttaaat gcggaattct accgggtagg ggaggcgctt
     2041 ttcccaaggc agtctggagc atgcgcttta gcagccccgc tggcacttgg cgctacacaa
     2101 gtggcctctg gcctcgcaca cattccacat ccaccggtag cgccaaccgg ctccgttctt
     2161 tggtggcccc ttcgcgccac cttctactcc tcccctagtc aggaagttcc cccccgcccc
     2221 gcagctcgcg tcgtgcagga cgtgacaaat ggaagtagca cgtctcacta gtctcgtgca
     2281 gatggacagc accgctgagc aatggaagcg ggtaggcctt tggggcagcg gccaatagca
     2341 gctttgctcc ttcgctttct gggctcagag gctgggaagg ggtgggtccg ggggcgggct
     2401 caggggcggg ctcaggggcg gggcgggcgc gaaggtcctc ccgaggcccg gcattctcgc
     2461 acgcttcaaa agcgcacgtc tgccgcgctg ttctcctctt cctcatctcc gggcctttcg
     2521 acctgcagcg acccgcttaa cagcgtcaac agcgtgccgc agatcttggt ggcgtgaaac
     2581 tcccgcacct cttcggccag cgccttgtag aagcgcgtat ggcttcgtac ccctgccatc
     2641 aacacgcgtc tgcgttcgac caggctgcgc gttctcgcgg ccataacaac cgacgtacgg
     2701 cgttgcgccc tcgccggcaa caaaaagcca cggaagtccg cctggagcag aaaatgccca
     2761 cgctactgcg ggtttatata gacggtcccc acgggatggg gaaaaccacc accacgcaac
     2821 tgctggtggc cctgggttcg cgcgacgata tcgtctacgt acccgagccg atgacttact
     2881 ggcgggtgtt gggggcttcc gagacaatcg cgaacatcta caccacacaa caccgcctcg
     2941 accagggtga gatatcggcc ggggacgcgg cggtggtaat gacaagcgcc cagataacaa
     3001 tgggcatgcc ttatgccgtg accgacgccg ttctggctcc tcatatcggg ggggaggctg
     3061 ggagctcaca tgccccgccc ccggccctca ccctcatctt cgaccgccat cccatcgccg
     3121 ccctcctgtg ctacccggcc gcgcgatacc ttatgggcag catgaccccc caggccgtgc
     3181 tggcgttcgt ggccctcatc ccgccgacct tgcccggcac aaacatcgtg ttgggggccc
     3241 ttccggagga cagacacatc gaccgcctgg ccaaacgcca gcgccccggc gagcggcttg
     3301 acctggctat gctggccgcg attcgccgcg tttatgggct gcttgccaat acggtgcggt
     3361 atctgcaggg cggcgggtcg tggcgggagg attggggaca gctttcgggg gcggccgtgc
     3421 cgccccaggg tgccgagccc cagagcaacg cgggcccacg accccatatc ggggacacgt
     3481 tatttaccct gtttcgggcc cccgagttgc tggcccccaa cggcgacctg tataacgtgt
     3541 ttgcctgggc tttggacgtc ttggccaaac gcctccgtcc catgcatgtc tttatcctgg
     3601 attacgacca atcgcccgcc ggctgccggg acgccctgct gcaacttacc tccgggatgg
     3661 tccagaccca cgtcaccacc ccaggctcca taccgacgat ctgcgacctg gcgcgcacgt
     3721 ttgcccggga gatgggggag gctaactgaa acacggaagg agacaatacc ggaaggaacc
     3781 cgcgctatga cggcaataaa aagacagaat aaaacgcacg ggtgttgggt cgtttgttca
     3841 taaacgcggg gttcggtccc agggctggca ctctgtcgat accccaccga gaccccattg
     3901 ggaccaatac gcccgcgttt cttccttttc cccaccccaa cccccaagtt cgggtgaagg
     3961 cccagggctc gcagccaacg tcggggcggc aagccctgcc atagccacgg gccccgtggg
     4021 ttagggacgg ggtcccccat ggggaatggt ttatggttcg tgggggttat tattttgggc
     4081 gttgcgtggg gtcaggtcca cgactggact gagcagacag acccatggtt tttggatggc
     4141 ctgggcatgg accgcatgta ctggcgcgac acgaacaccg ggcgtctgtg gctgccaaac
     4201 acccccgacc cccaaaaacc accgcgcgga tttctggcgc cgccggacga actaaacctg
     4261 actacggcat ctctgcccct tcttcgctgg tacgaggagc gcttttgttt tgtattggtc
     4321 accacggccg agtttccgcg ggaccccggc caggacctgc agaaattgat gatctattaa
     4381 acaataaaga tgtccactaa aatggaagtt tttcctgtca tactttgtta agaagggtga
     4441 gaacagagta cctacatttt gaatggaagg attggagcta cgggggtggg ggtggggtgg
     4501 gattagataa atgcctgctc tttactgaag gctctttact attgctttat gataatgttt
     4561 catagttgga tatcataatt taaacaagca aaaccaaatt aagggccagc tcattcctcc
     4621 cactcatgat ctatagatct atagatctct cgtgggatca ttgtttttct cttgattccc
     4681 actttgtggt tctaagtact gtggtttcca aatgtgtcag tttcatagcc tgaagaacga
     4741 gatcagcagc ctctgttcca catacacttc attctcagta ttgttttgcc aagttctaat
     4801 tccatcagaa gcttggcact ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc
     4861 gttacccaac ttaatcgcct tgcagcacat ccccctttcg ccagctggcg taatagcgaa
     4921 gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga atggcgcctg
     4981 atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatatg gtgcactctc
     5041 agtacaatct gctctgatgc cgcatagtta agccagcccc gacacccgcc aacacccgct
     5101 gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc
     5161 tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag
     5221 ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg
     5281 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     5341 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     5401 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
     5461 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
     5521 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
     5581 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
     5641 gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct
     5701 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
     5761 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
     5821 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
     5881 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
     5941 gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta
     6001 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
     6061 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
     6121 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
     6181 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     6241 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
     6301 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     6361 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     6421 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     6481 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     6541 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg
     6601 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     6661 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     6721 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     6781 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     6841 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     6901 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     6961 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     7021 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     7081 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
     7141 tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc
     7201 gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat
     7261 taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt
     7321 aatgtgagtt agctcactca ttaggcaccc caggctttac actttatgct tccggctcgt
     7381 atgttgtgtg gaattgtgag cggataacaa tttcacacag gaaacagcta tgaccatgat
     7441 tacgccaagc tcgaaattaa ccctcactaa agggaacaaa agctggagct ccaccgcggt
     7501 g
//
