LOCUS       sequence-388198-        7937 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For loading Cre into PVCs; also contains PVC regulatory genes .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7937)
  AUTHORS   Kreitz J, Friedrich MJ, Guru A, Lash B, Saito M, Macrae RK, Zhang F
  TITLE     Programmable protein delivery with a bacterial contractile injection
            system.
  JOURNAL   Nature. 2023 Apr;616(7956):357-364. doi: 10.1038/s41586-023-05870-7.
            Epub 2023 Mar 29.
  PUBMED    36991127
REFERENCE   2  (bases 1 to 7937)
  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"; doi: 
            "10.1038/s41586-023-05870-7"; journalName: "Nature"; date: 
            "2023-04"; volume: "616"; issue: "7956"; pages: "357-364"
COMMENT     Sequence Label: pBR322-Pdp1_NTD-Cre
FEATURES             Location/Qualifiers
     source          1..7937
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             3463..3654
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(3648..3670)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    3756..3896
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     3811..3830
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(3911..3928)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4082..4670)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4162..4181)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4841..5632,5633..5701))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     5464..5483
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5702..5806)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5874..5892
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(5919..5924,5925..5941,5942..5947)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     CDS             6861..6884
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             6888..6908
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6909..7937
                     /codon_start=1
                     /product="site-specific recombinase"
                     /label=Cre
                     /note="Cre is a site-specific recombinase from 
                     bacteriophage P1. Recombination occurs at loxP sequences."
                     /translation="SNLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAFSEHTWKMLQ
                     SVCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNMLHRRSGLP
                     RPSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRCQDIRNLAFL
                     GIAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALSLGVTKLVER
                     WISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYGAKDDSGQR
                     YLAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRNLDSETGAMVRLLED
                     GD"
ORIGIN
        1 tatggatttt catgtcatca ggagaatatt caaaattctg tgtcacacaa aatattttac
       61 agcgtaatca caccattaag aggggcatca aaataaatgg ttaactggga caatgttgaa
      121 tttcattttt ataaggcatt gtccgtatta tccaccatcc agatcaacgg attcagatgg
      181 tgtaaagaaa gcatatagct ggtaggtaaa attaagaagt gtagttttat aaagtattaa
      241 tggagaacga tgaatttaaa gtaaatggag gaaggttagt ttcactatca aattacagga
      301 ccaaaaatat ggagtattcc atcttctttg gtgaattaag tcttgaagac ttcaacaaat
      361 attgcctgga actatgggcc tgtgaatggt attggagtat catttataac gactatttcg
      421 ataggaaatt taaagtcttg ttgaaaatga ctaatcgtct aaaagtatgt ctggagaaat
      481 aggtttaatc tctatattta acggttttat tttcatctat ttactaatgg taaatttttt
      541 tactctttga atatattggt gagtttgtta tccccattag ttaccagggt attattaatg
      601 agtgaaattc aattttattg atattaaagt caaatttaca aatgatgact ttattatttt
      661 ttatttatta attttaaatt attttttgtt aatctttact ataaatgaat tttttgcttg
      721 catggacttg atttttttat atataacatt gacaaatatt aattcatata tatgattgtt
      781 atagtgagtt atttaattga ttcgactaac aatttcattg taatttttca agatagcagt
      841 caattgctat aacaagcatc tttctcgtca aaggaaaatg tgtatggcct ataagcagca
      901 acttgaaaga tgaaggatat aatgatgatt aacttaacgg agagtagata ttagaagtag
      961 tagaaggaca atttaagact ttatacccat taaaacaaag gggtaaaata atgatgtttg
     1021 atgaaaattt agaatgtgtt aatcacgaaa ttatcaatga aaaattagat acgaaaattc
     1081 accttgctaa tccttatgcc ttttgggggg gaagggtaaa tgagaatata aatgggttaa
     1141 ttaaacaaca ttctttaaga ggaactgact ttaataaaat ttttaaccgg aagataaatt
     1201 tttttgtaaa tggaataaat aattgccccc gaaaaactcg gaatataaaa ccccctaatt
     1261 aattatttaa aggaataaga gtttatttgc tcgtagagta attatgttcc attaatatgc
     1321 taaattgctc agtaaagtta ttaacttttt ttcaacgcca gtgaataagt ctgttttatt
     1381 aaataaggag ttcaggtgtt tatttcaaaa gaattatcaa gctttattgc tgtagctaaa
     1441 aataaatcaa taaataaagc aacaacggaa ctttttatta cacagtcacc gatttgccga
     1501 agcttaaaaa aactggaatt ttcattaggt gtaaaactct ttgtaagaaa aagccaggga
     1561 ttggctttaa cagaagaagg agaatgctta tataataaag ttttgtcaat ttgtgaacgg
     1621 cttcaagaaa tagaaggtga ctacaagaat gaaaaatcaa catacaatga tctatatcaa
     1681 aaaaatatat gaggttctaa tataataagc ggttctagat aactaaagtc atctattacc
     1741 aatatctgta tactcagtga aagttttgtc ttctaggtaa gttttcataa tagtgacata
     1801 ccttattttt tcatctgggt gtctctcatc ctgaagaatg ggtgatatag ttgaaataac
     1861 tcagttcgtt ttgtggttta cgatttttgg cggatcacaa aaatagggct gagtcccttc
     1921 aagtgagttt tgaaatatac aattataccc ttcatccttt aagttgttgc tttgttggct
     1981 gttttcattc acctcagtca catagttatc tatgctcctg aggattcgtt cacttgccgt
     2041 cgtgctgcaa ctcgaaatct attgggtata tatgtaattt tcatattctg tttcatttta
     2101 acggtattat ttaaatattt atatttagaa cattaggtta ttagaataga taattttagg
     2161 taaatgtaat taattcaacc atcacggaga gtttatcagt cgaccgatgc ccttgagagc
     2221 cttcaaccca gtcagctcct tccggtgggc gcggggcatg actatcgtcg ccgcacttat
     2281 gactgtcttc tttatcatgc aactcgtagg acaggtgccg gcagcgctct gggtcatttt
     2341 cggcgaggac cgctttcgct ggagcgcgac gatgatcggc ctgtcgcttg cggtattcgg
     2401 aatcttgcac gccctcgctc aagccttcgt cactggtccc gccaccaaac gtttcggcga
     2461 gaagcaggcc attatcgccg gcatggcggc cgacgcgctg ggctacgtct tgctggcgtt
     2521 cgcgacgcga ggctggatgg ccttccccat tatgattctt ctcgcttccg gcggcatcgg
     2581 gatgcccgcg ttgcaggcca tgctgtccag gcaggtagat gacgaccatc agggacagct
     2641 tcaaggatcg ctcgcggctc ttaccagcct aacttcgatc attggaccgc tgatcgtcac
     2701 ggcgatttat gccgcctcgg cgagcacatg gaacgggttg gcatggattg taggcgccgc
     2761 cctatacctt gtctgcctcc ccgcgttgcg tcgcggtgca tggagccggg ccacctcgac
     2821 ctgaatggaa gccggcggca cctcgctaac ggattcacca ctccaagaat tggagccaat
     2881 caattcttgc ggagaactgt gaatgcgcaa accaaccctt ggcagaacat atccatcgcg
     2941 tccgccatct ccagcagccg cacgcggcgc atctcgggca gcgttgggtc ctggccacgg
     3001 gtgcgcatga tcgtgctcct gtcgttgagg acccggctag gctggcgggg ttgccttact
     3061 ggttagcaga atgaatcacc gatacgcgag cgaacgtgaa gcgactgctg ctgcaaaacg
     3121 tctgcgacct gagcaacaac atgaatggtc ttcggtttcc gtgtttcgta aagtctggaa
     3181 acgcggaagt cagcgccctg caccattatg ttccggatct gcatcgcagg atgctgctgg
     3241 ctaccctgtg gaacacctac atctgtatta acgaagcgct ggcattgacc ctgagtgatt
     3301 tttctctggt cccgccgcat ccataccgcc agttgtttac cctcacaacg ttccagtaac
     3361 cgggcatgtt catcatcagt aacccgtatc gtgagcatcc tctctcgttt catcggtatc
     3421 attaccccca tgaacagaaa tcccccttac acggaggcat cagtgaccaa acaggaaaaa
     3481 accgccctta acatggcccg ctttatcaga agccagacat taacgcttct ggagaaactc
     3541 aacgagctgg acgcggatga acaggcagac atctgtgaat cgcttcacga ccacgctgat
     3601 gagctttacc gcagctgcct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg
     3661 cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt
     3721 cagggcgcgt cagcgggtgt tggcgggtgt cggggcgcag ccatgaccca gtcacgtagc
     3781 gatagcggag tgtatactgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
     3841 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgct
     3901 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     3961 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     4021 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     4081 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     4141 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     4201 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     4261 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     4321 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     4381 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     4441 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     4501 ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta
     4561 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     4621 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     4681 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     4741 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     4801 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     4861 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     4921 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     4981 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     5041 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     5101 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctgcag
     5161 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     5221 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     5281 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     5341 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     5401 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaacac
     5461 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     5521 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     5581 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     5641 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     5701 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     5761 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     5821 aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc
     5881 gtatcacgag gccctttcgt cttcaagaat tctcatgttt gacagcttat catcgataag
     5941 ctttaatgcg gtagtttatc acagttaaat tgctaacgca gtcaggcacc gtgtatgaaa
     6001 tctaacaatg cgctcatcgt catcctcggc accgtcaccc tggatgctgt aggcataggc
     6061 ttggttatgc cggtactgcc gggcctcttg cgggatatcg tccattccga cagcatcgcc
     6121 agtcactatg gcgtgctgct agcgctatat gcgttgatgc aatttctatg cgcacccgtt
     6181 ctcggagcac tgtccgaccg ctttggccgc cgcccagtcc tgctcgcttc gctacttgga
     6241 gccactatcg actacgcgat catggcgacc acacccgtcc tgtggatcca tttttttaat
     6301 taaataaagg aattaatatt gatttgatag agcatgtaat caataacttc tatgaaaatg
     6361 ttgaaaaaca atacacggaa acacttaact tacaattaaa ttgttaaagg agatgatgat
     6421 aagaatggtc agtcaccaga taaaatgatc tctggtcaat tttaggaaag ctgaaaaaac
     6481 ctgaccaaat gcgtcgtaaa ttctatgctt agaaagactc ggtgccagtg taatacctaa
     6541 gatatggttt ttatttccat atctgttcga aaacttagaa ataacaagtt caacaataac
     6601 cagggacttt aaatatgcct agatatgcta attatcagat aaaccccaaa cagaatatta
     6661 aaaattcaca tgggaaatct tcatcgtcag atttttctag tgggtacctt tcattctcaa
     6721 ataattcgct tgatgaccct tttattcggc agcaagttaa gagagaattt atttgggagg
     6781 gacatatgaa ggagattgaa gaagcttcaa gattaggtgg ctctggtggt ggcggttctg
     6841 gcggcggtgg ttctggcggt gattataagg acgatgacga taaggatccc aagaaaaagc
     6901 gcaaggtatc caatttactg accgtacacc aaaatttgcc tgcattaccg gtcgatgcaa
     6961 cgagtgatga ggttcgcaag aacctgatgg acatgttcag ggatcgccag gcgttttctg
     7021 agcatacctg gaaaatgctt cagtccgttt gccggtcgtg ggcggcatgg tgcaagttga
     7081 ataaccggaa atggtttccc gcagaacctg aagatgttcg cgattatctt ctatatcttc
     7141 aggcgcgcgg tctggcagta aaaactatcc agcaacattt gggccagcta aacatgcttc
     7201 atcgtcggtc cgggctgcca cgaccaagtg acagcaatgc tgtttcactg gttatgcggc
     7261 ggatccgaaa agaaaacgtt gatgccggtg aacgtgcaaa acaggctcta gcgttcgaac
     7321 gcactgattt cgaccaggtt cgttcactca tggaaaatag cgatcgctgc caggatatac
     7381 gtaatctggc atttctgggg attgcttata acaccctgtt acgtatagcc gaaattgcca
     7441 ggatcagggt taaagatatc tcacgtactg acggtgggag aatgttaatc catattggca
     7501 gaacgaaaac gctggttagc accgctggtg tagagaaggc acttagcctg ggggtaacta
     7561 aactggtcga gcgatggatt tccgtctctg gtgtagctga tgatccgaat aactacctgt
     7621 tttgccgggt cagaaaaaat ggtgttgccg cgccatctgc caccagccag ctatcaactc
     7681 gcgccctgga agggattttt gaagcaactc atcgattgat ttacggcgct aaggatgact
     7741 ctggtcagag atacctggcc tggtctggac acagtgcccg tgtcggagcc gcgcgagata
     7801 tggcccgcgc tggagtttca ataccggaga tcatgcaagc tggtggctgg accaatgtaa
     7861 atattgtcat gaactatatc cgtaacctgg atagtgaaac aggggcaatg gtgcgcctgc
     7921 tggaagatgg cgattaa
//
