LOCUS       sequence-388358-        9453 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Loads HiBiT-tagged Cre into PVCs using Pdp1 packaging domain; also 
            contains Pnf .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9453)
  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 9453)
  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-HiBiT
FEATURES             Location/Qualifiers
     source          1..9453
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             3214..3405
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(3399..3421)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    3507..3647
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     3562..3581
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(3662..3679)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3833..4421)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3913..3932)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4592..5383,5384..5452))
                     /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     5215..5234
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5453..5557)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5625..5643
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(5670..5675,5676..5692,5693..5698)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     CDS             6612..6635
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             6639..6659
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6660..7685
                     /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 aactaaagtc atctattacc aatatctgta tactcagtga aagttttgtc ttctaggtaa
       61 gttttcataa tagtgacata ccttattttt tcatctgggt gtctctcatc ctgaagaatg
      121 ggtgatatag ttgaaataac tcagttcgtt ttgtggttta cgatttttgg cggatcacaa
      181 aaatagggct gagtcccttc aagtgagttt tgaaatatac aattataccc ttcatccttt
      241 aagttgttgc tttgttggct gttttcattc acctcagtca catagttatc tatgctcctg
      301 aggattcgtt cacttgccgt cgtgctgcaa ctcgaaatct attgggtata tatgtaattt
      361 tcatattctg tttcatttta acggtattat ttaaatattt atatttagaa cattaggtta
      421 ttagaataga taattttagg taaatgtaat taattcaacc atcacggaga gtttatcaat
      481 gttaaaatat gctaatcctc agaccgtagc cacacaacgt actaaaaata ctgcgaagaa
      541 accgccatca tcaacctctt ttgatgggca ccttgaactt tcaaatggtg aaaaccagcc
      601 ttacgaaggc cataagatta ggaaaatcaa aggattgaga caacatcttg cggataggag
      661 tcttaataaa gggcatattt ccccgttgat gaataaggga ttattagtag gtagtaagga
      721 tgtgtcaatt gacatcccag ttatagctca tcgttatgac agcagccatc aacttacaga
      781 tgcggagcct ttgaaagctg atagtcactc caatcatcta gacccttttt atggtgttat
      841 tgcaggattt cgtggtgatc aggtgacatc atcagagtcg ggttcaggat caataggcgt
      901 acattggggg aaaaatacgc tggattccaa tattatgggt gttaatgtgg ttaatggtgc
      961 atcgggaacc gtgggaataa gaattgcttt gaaggatatt cagcatgggt ccccggttat
     1021 tgtcacgtct ggagcattga gcggttgtac tatggtatat tcggtgaaaa atggctattt
     1081 ttttgcctat catacggggc agaaaccggg taataatgaa tggaaaacag gcaggcaggg
     1141 tgttgttgct acctatctga gccatcaagc cctttcgccg gatagtgaac ctatgacggt
     1201 tggtgaacag aataacgatt tagtgaatat ttttgcaaat tatgatcaga gtgttattac
     1261 ctatatggga aaaccgggag tgctgattga taaaatggca gaaaatgttg gtgtttttaa
     1321 ttatgacgaa attaagcctg aaaaacctgc tattagagct ggctacagtt atgcgttgct
     1381 ggctaaggac gataaaggaa aggtgaacgt taaggtacta tcggaagatg ttatcgtttc
     1441 atcaggtaag cagggaaata ctgttaaggc cattaattca cttaaaaaac ggttgttata
     1501 attttgccca atgagggcag gaggtacgga ttttcattag cacttatcga agcggatata
     1561 tgtttgtgac tacattgtca gaaaatttca ttcatcaact tgaattagaa cagtgtccat
     1621 ataacttatt tagaataaag gaaggaaatt aatcttagat acaatgataa gggaaataaa
     1681 atagaaagta gagttaattg tattaataag gtgctggagc tttacttaca aaatatcaat
     1741 gatttttggt ttcgtataag taattaattt agagtaatga aaaattattt gtgggttaaa
     1801 ttttatttat tgaaaattca ttggtttatt ttatgaaata ggaaggtaat agcggtatta
     1861 ttcggttgaa ataaccgtat agataaatag taggatttag ataaattatt tcatatatga
     1921 caatttttac tattatatgg ggtttgtaag tcgaccgatg cccttgagag ccttcaaccc
     1981 agtcagctcc ttccggtggg cgcggggcat gactatcgtc gccgcactta tgactgtctt
     2041 ctttatcatg caactcgtag gacaggtgcc ggcagcgctc tgggtcattt tcggcgagga
     2101 ccgctttcgc tggagcgcga cgatgatcgg cctgtcgctt gcggtattcg gaatcttgca
     2161 cgccctcgct caagccttcg tcactggtcc cgccaccaaa cgtttcggcg agaagcaggc
     2221 cattatcgcc ggcatggcgg ccgacgcgct gggctacgtc ttgctggcgt tcgcgacgcg
     2281 aggctggatg gccttcccca ttatgattct tctcgcttcc ggcggcatcg ggatgcccgc
     2341 gttgcaggcc atgctgtcca ggcaggtaga tgacgaccat cagggacagc ttcaaggatc
     2401 gctcgcggct cttaccagcc taacttcgat cattggaccg ctgatcgtca cggcgattta
     2461 tgccgcctcg gcgagcacat ggaacgggtt ggcatggatt gtaggcgccg ccctatacct
     2521 tgtctgcctc cccgcgttgc gtcgcggtgc atggagccgg gccacctcga cctgaatgga
     2581 agccggcggc acctcgctaa cggattcacc actccaagaa ttggagccaa tcaattcttg
     2641 cggagaactg tgaatgcgca aaccaaccct tggcagaaca tatccatcgc gtccgccatc
     2701 tccagcagcc gcacgcggcg catctcgggc agcgttgggt cctggccacg ggtgcgcatg
     2761 atcgtgctcc tgtcgttgag gacccggcta ggctggcggg gttgccttac tggttagcag
     2821 aatgaatcac cgatacgcga gcgaacgtga agcgactgct gctgcaaaac gtctgcgacc
     2881 tgagcaacaa catgaatggt cttcggtttc cgtgtttcgt aaagtctgga aacgcggaag
     2941 tcagcgccct gcaccattat gttccggatc tgcatcgcag gatgctgctg gctaccctgt
     3001 ggaacaccta catctgtatt aacgaagcgc tggcattgac cctgagtgat ttttctctgg
     3061 tcccgccgca tccataccgc cagttgttta ccctcacaac gttccagtaa ccgggcatgt
     3121 tcatcatcag taacccgtat cgtgagcatc ctctctcgtt tcatcggtat cattaccccc
     3181 atgaacagaa atccccctta cacggaggca tcagtgacca aacaggaaaa aaccgccctt
     3241 aacatggccc gctttatcag aagccagaca ttaacgcttc tggagaaact caacgagctg
     3301 gacgcggatg aacaggcaga catctgtgaa tcgcttcacg accacgctga tgagctttac
     3361 cgcagctgcc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg
     3421 gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg
     3481 tcagcgggtg ttggcgggtg tcggggcgca gccatgaccc agtcacgtag cgatagcgga
     3541 gtgtatactg gcttaactat gcggcatcag agcagattgt actgagagtg caccatatgc
     3601 ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggcgc tcttccgctt
     3661 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     3721 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     3781 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     3841 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     3901 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     3961 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     4021 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     4081 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     4141 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     4201 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     4261 gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     4321 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagctgt ggtttttttg
     4381 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     4441 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     4501 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     4561 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     4621 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     4681 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     4741 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     4801 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     4861 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctgca ggcatcgtgg
     4921 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     4981 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     5041 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     5101 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     5161 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaaca cgggataata
     5221 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     5281 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     5341 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     5401 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     5461 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     5521 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     5581 ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga
     5641 ggccctttcg tcttcaagaa ttctcatgtt tgacagctta tcatcgataa gctttaatgc
     5701 ggtagtttat cacagttaaa ttgctaacgc agtcaggcac cgtgtatgaa atctaacaat
     5761 gcgctcatcg tcatcctcgg caccgtcacc ctggatgctg taggcatagg cttggttatg
     5821 ccggtactgc cgggcctctt gcgggatatc gtccattccg acagcatcgc cagtcactat
     5881 ggcgtgctgc tagcgctata tgcgttgatg caatttctat gcgcacccgt tctcggagca
     5941 ctgtccgacc gctttggccg ccgcccagtc ctgctcgctt cgctacttgg agccactatc
     6001 gactacgcga tcatggcgac cacacccgtc ctgtggatcc atttttttaa ttaaataaag
     6061 gaattaatat tgatttgata gagcatgtaa tcaataactt ctatgaaaat gttgaaaaac
     6121 aatacacgga aacacttaac ttacaattaa attgttaaag gagatgatga taagaatggt
     6181 cagtcaccag ataaaatgat ctctggtcaa ttttaggaaa gctgaaaaaa cctgaccaaa
     6241 tgcgtcgtaa attctatgct tagaaagact cggtgccagt gtaataccta agatatggtt
     6301 tttatttcca tatctgttcg aaaacttaga aataataagt tcaacaataa ccagggactt
     6361 taaatatgcc tagatatgct aattatcaga taaaccccaa acagaatatt aaaaattcac
     6421 atgggaaatc ttcatcgtca gatttttcta gtgggtacct ttcattctca aataattcgc
     6481 ttgatgaccc ttttattcgg cagcaagtta agagagaatt tatttgggag ggacatatga
     6541 aggagattga agaagcttca agattaggtg gctctggtgg tggcggttct ggcggcggtg
     6601 gttctggcgg tgattataag gacgatgacg ataaggatcc caagaaaaag cgcaaggtat
     6661 ccaatttact gaccgtacac caaaatttgc ctgcattacc ggtcgatgca acgagtgatg
     6721 aggttcgcaa gaacctgatg gacatgttca gggatcgcca ggcgttttct gagcatacct
     6781 ggaaaatgct tcagtccgtt tgccggtcgt gggcggcatg gtgcaagttg aataaccgga
     6841 aatggtttcc cgcagaacct gaagatgttc gcgattatct tctatatctt caggcgcgcg
     6901 gtctggcagt aaaaactatc cagcaacatt tgggccagct aaacatgctt catcgtcggt
     6961 ccgggctgcc acgaccaagt gacagcaatg ctgtttcact ggttatgcgg cggatccgaa
     7021 aagaaaacgt tgatgccggt gaacgtgcaa aacaggctct agcgttcgaa cgcactgatt
     7081 tcgaccaggt tcgttcactc atggaaaata gcgatcgctg ccaggatata cgtaatctgg
     7141 catttctggg gattgcttat aacaccctgt tacgtatagc cgaaattgcc aggatcaggg
     7201 ttaaagatat ctcacgtact gacggtggga gaatgttaat ccatattggc agaacgaaaa
     7261 cgctggttag caccgctggt gtagagaagg cacttagcct gggggtaact aaactggtcg
     7321 agcgatggat ttccgtctct ggtgtagctg atgatccgaa taactacctg ttttgccggg
     7381 tcagaaaaaa tggtgttgcc gcgccatctg ccaccagcca gctatcaact cgcgccctgg
     7441 aagggatttt tgaagcaact catcgattga tttacggcgc taaggatgac tctggtcaga
     7501 gatacctggc ctggtctgga cacagtgccc gtgtcggagc cgcgcgagat atggcccgcg
     7561 ctggagtttc aataccggag atcatgcaag ctggtggctg gaccaatgta aatattgtca
     7621 tgaactatat ccgtaacctg gatagtgaaa caggggcaat ggtgcgcctg ctggaagatg
     7681 gcgatgggag ctccggtgtg agcggctggc ggctgttcaa gaagattagc taatatggat
     7741 tttcatgtca tcaggagaat attcaaaatt ctgtgtcaca caaaatattt tacagcgtaa
     7801 tcacaccatt aagaggggca tcaaaataaa tggttaactg ggacaatgtt gaatttcatt
     7861 tttataaggc attgtccgta ttatccacca tccagatcaa cggattcaga tggtgtaaag
     7921 aaagcatata gctggtaggt aaaattaaga agtgtagttt tataaagtat taatggagaa
     7981 cgatgaattt aaagtaaatg gaggaaggtt agtttcacta tcaaattaca ggaccaaaaa
     8041 tatggagtat tccatcttct ttggtgaatt aagtcttgaa gacttcaaca aatattgcct
     8101 ggaactatgg gcctgtgaat ggtattggag tatcatttat aacgactatt tcgataggaa
     8161 atttaaagtc ttgttgaaaa tgactaatcg tctaaaagta tgtctggaga aataggttta
     8221 atctctatat ttaacggttt tattttcatc tatttactaa tggtaaattt ttttactctt
     8281 tgaatatatt ggtgagtttg ttatccccat tagttaccag ggtattatta atgagtgaaa
     8341 ttcaatttta ttgatattaa agtcaaattt acaaatgatg actttattat tttttattta
     8401 ttaattttaa attatttttt gttaatcttt actataaatg aattttttgc ttgcatggac
     8461 ttgatttttt tatatataac attgacaaat attaattcat atatatgatt gttatagtga
     8521 gttatttaat tgattcgact aacaatttca ttgtaatttt tcaagatagc agtcaattgc
     8581 tataacaagc atctttctcg tcaaaggaaa atgtgtatgg cctataagca gcaacttgaa
     8641 agatgaagga tataatgatg attaacttaa cggagagtag atattagaag tagtagaagg
     8701 acaatttaag actttatacc cattaaaaca aaggggtaaa ataatgatgt ttgatgaaaa
     8761 tttagaatgt gttaatcacg aaattatcaa tgaaaaatta gatacgaaaa ttcaccttgc
     8821 taatccttat gccttttggg ggggaagggt aaatgagaat ataaatgggt taattaaaca
     8881 acattcttta agaggaactg actttaataa aatttttaac cggaagataa atttttttgt
     8941 aaatggaata aataattgcc cccgaaaaac tcggaatata aaacccccta attaattatt
     9001 taaaggaata agagtttatt tgctcgtaga gtaattatgt tccattaata tgctaaattg
     9061 ctcagtaaag ttattaactt tttttcaacg ccagtgaata agtctgtttt attaaataag
     9121 gagttcaggt gtttatttca aaagaattat caagctttat tgctgtagct aaaaataaat
     9181 caataaataa agcaacaacg gaacttttta ttacacagtc accgatttgc cgaagcttaa
     9241 aaaaactgga attttcatta ggtgtaaaac tctttgtaag aaaaagccag ggattggctt
     9301 taacagaaga aggagaatgc ttatataata aagttttgtc aatttgtgaa cggcttcaag
     9361 aaatagaagg tgactacaag aatgaaaaat caacatacaa tgatctatat caaaaaaata
     9421 tatgaggttc taatataata agcggttcta gat
//
