LOCUS       sequence-388716-        4704 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Cre in insect cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4704)
  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 4704)
  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: pIEx4-Cre
FEATURES             Location/Qualifiers
     source          1..4704
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             4..27
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             31..51
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             52..1080
                     /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"
     CDS             1095..1118
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     terminator      1137..1444
                     /label=IE1 terminator
                     /note="terminator of the ie1 gene from the baculovirus 
                     Autographa californica"
     promoter        1474..1578
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1579..1647,1648..2439)
                     /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(1797..1816)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2610..3198
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3099..3118
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3352..3369
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3486..3507
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3522..3527,3528..3545,3546..3552)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3560..3576
                     /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     3565..3587
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3584..3600
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3584..3600
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        3621..4103
                     /gene="Autographa californica hr5"
                     /label=hr5 enhancer
                     /note="baculovirus early transcription enhancer"
     promoter        4107..4698
                     /label=IE1 promoter
                     /note="promoter of the ie1 gene from the baculovirus 
                     Autographa californica"
     regulatory      join(4699..4704,1..4)
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 atggattata aggacgatga cgataaggat cccaagaaaa agcgcaaggt atccaattta
       61 ctgaccgtac accaaaattt gcctgcatta ccggtcgatg caacgagtga tgaggttcgc
      121 aagaacctga tggacatgtt cagggatcgc caggcgtttt ctgagcatac ctggaaaatg
      181 cttcagtccg tttgccggtc gtgggcggca tggtgcaagt tgaataaccg gaaatggttt
      241 cccgcagaac ctgaagatgt tcgcgattat cttctatatc ttcaggcgcg cggtctggca
      301 gtaaaaacta tccagcaaca tttgggccag ctaaacatgc ttcatcgtcg gtccgggctg
      361 ccacgaccaa gtgacagcaa tgctgtttca ctggttatgc ggcggatccg aaaagaaaac
      421 gttgatgccg gtgaacgtgc aaaacaggct ctagcgttcg aacgcactga tttcgaccag
      481 gttcgttcac tcatggaaaa tagcgatcgc tgccaggata tacgtaatct ggcatttctg
      541 gggattgctt ataacaccct gttacgtata gccgaaattg ccaggatcag ggttaaagat
      601 atctcacgta ctgacggtgg gagaatgtta atccatattg gcagaacgaa aacgctggtt
      661 agcaccgctg gtgtagagaa ggcacttagc ctgggggtaa ctaaactggt cgagcgatgg
      721 atttccgtct ctggtgtagc tgatgatccg aataactacc tgttttgccg ggtcagaaaa
      781 aatggtgttg ccgcgccatc tgccaccagc cagctatcaa ctcgcgccct ggaagggatt
      841 tttgaagcaa ctcatcgatt gatttacggc gctaaggatg actctggtca gagatacctg
      901 gcctggtctg gacacagtgc ccgtgtcgga gccgcgcgag atatggcccg cgctggagtt
      961 tcaataccgg agatcatgca agctggtggc tggaccaatg taaatattgt catgaactat
     1021 atccgtaacc tggatagtga aacaggggca atggtgcgcc tgctggaaga tggcgattaa
     1081 cttctggcct cgagcaccac catcaccatc accatcacta agtgattaac ctcaggttat
     1141 acatatattt tgaatttaat taattataca tatattttat attatttttg tcttttatta
     1201 tcgaggggcc gttgttggtg tggggttttg catagaaata acaatgggag ttggcgacgt
     1261 tgctgcgcca acaccacctc ccttccctcc tttcatcatg tatctgtaga taaaataaaa
     1321 tattaaacct aaaaacaaga ccgcgcctat caacaaaatg ataggcatta acttgccgct
     1381 gacgctgtca ctaacgttgg acgatttgcc gactaaacct tcatcgccca gtaaccaatc
     1441 tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     1501 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     1561 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
     1621 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     1681 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     1741 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
     1801 tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac
     1861 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     1921 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     1981 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     2041 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     2101 gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc
     2161 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     2221 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     2281 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     2341 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     2401 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
     2461 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     2521 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     2581 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     2641 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     2701 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     2761 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     2821 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     2881 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     2941 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     3001 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     3061 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     3121 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     3181 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     3241 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     3301 accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca
     3361 gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg
     3421 attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac
     3481 gcaattaatg tgagttagct cactcattag gcaccccagg ctttacactt tatgcttccg
     3541 gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagctatgac
     3601 catgattacg aattcccggg cgcgtaaaac acaatcaagt atgagtcata agctgatgtc
     3661 atgttttgca cacggctcat aaccgaactg gctttacgag tagaattcta cttgtaacgc
     3721 acgatcagtg gatgatgtca tttgtttttc aaatcgagat gatgtcatgt tttgcacacg
     3781 gctcataaac tcgctttacg agtagaattc tacgtgtaac gcacgatcga ttgatgagtc
     3841 atttgttttg caatatgata tcatacaata tgactcattt gtttttcaaa accgaacttg
     3901 atttacgggt agaattctac ttgtaaagca caatcaaaaa gatgatgtca tttgtttttc
     3961 aaaactgaac tcgctttacg agtagaattc tacgtgtaaa acacaatcaa gaaatgatgt
     4021 catttgttat aaaaataaaa gctgatgtca tgttttgcac atggctcata actaaactcg
     4081 ctttacgggt agaattctac gcgcgtcgat gtctttgtga tgcgcgcgac atttttgtag
     4141 gttattgata aaatgaacgg atacgttgcc cgacattatc attaaatcct tggcgtagaa
     4201 tttgtcgggt ccattgtccg tgtgcgctag catgcccgta acggacctcg tacttttggc
     4261 ttcaaaggtt ttgcgcacag acaaaatgtg ccacacttgc agctctgcat gtgtgcgcgt
     4321 taccacaaat cccaacggcg cagtgtactt gttgtatgca aataaatctc gataaaggcg
     4381 cggcgcgcga atgcagctga tcacgtacgc tcctcgtgtt ccgttcaagg acggtgttat
     4441 cgacctcaga ttaatgttta tcggccgact gttttcgtat ccgctcacca aacgcgtttt
     4501 tgcattaaca ttgtatgtcg gcggatgttc tatatctaat ttgaataaat aaacgataac
     4561 cgcgttggtt ttagagggca taataaaaga aatattgtta tcgtgttcgc cattagggca
     4621 gtataaattg acgttcatgt tggatattgt ttcagttgca agttgacact ggcggcgaca
     4681 agatcgtgaa caaccaaggc cacc
//
