LOCUS       sequence-6971-ex        4870 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Cre recombinase from mouse PGK promoter in mammalian 
            cells..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4870)
  TITLE     Rajewsky pPGK-Cre-bpA
REFERENCE   2  (bases 1 to 4870)
  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: pPGK-Cre-bpA
FEATURES             Location/Qualifiers
     source          1..4870
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        52..156
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(157..225,226..1017)
                     /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(375..394)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1188..1776
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1677..1696
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1930..1947
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2064..2085
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2100..2105,2106..2123,2124..2130)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2138..2154
                     /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     2143..2165
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2162..2178
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2162..2178
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     2197..2217
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        2199..2217
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     2246..2262
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     2247..2263
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2323..2346)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     2712..2731
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             2804..2824
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2825..3853
                     /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="SNLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAFSEHTWKMLL
                     SVCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNMLHRRSGLP
                     RPSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRCQDIRNLAFL
                     GIAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALSLGVTKLVER
                     WISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYGAKDDSGQR
                     YLAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRNLDSETGAMVRLLED
                     GD"
     primer_bind     complement(3877..3893)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3907..3924)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3913..4137
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4228..4247)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4229..4247)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4257..4274)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4257..4273)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4266..4288)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4415..4843
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4502..4521)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4712..4733
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 atgcaatcct actatgcaat gtatccaggt ggcacttttc ggggaaatgt gcgcggaacc
       61 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
      121 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
      181 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
      241 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
      301 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
      361 acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa
      421 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
      481 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
      541 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
      601 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
      661 gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg
      721 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
      781 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
      841 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg
      901 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
      961 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
     1021 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     1081 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     1141 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     1201 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     1261 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     1321 ataccaaata ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     1381 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     1441 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     1501 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     1561 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     1621 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     1681 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     1741 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     1801 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     1861 tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg
     1921 accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct
     1981 ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa
     2041 gcgggcagtg agcgcaacgc aattaatgtg agttagctca ctcattaggc accccaggct
     2101 ttacacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata acaatttcac
     2161 acaggaaaca gctatgacca tgattacgcc aagcgcgcaa ttaaccctca ctaaagggaa
     2221 caaaagctgg gtaccgggcc ccccctcgag gtcgacggta tcgataagct tgatatcgaa
     2281 ttctaccggg taggggaggc gcttttccca aggcagtctg gagcatgcgc tttagcagcc
     2341 ccgctggcac ttggcgctac acaagtggcc tctggcctcg cacacattcc acatccaccg
     2401 gtagcgccaa ccggctccgt tctttggtgg ccccttcgcg ccacttctac tcctccccta
     2461 gtcaggaagt ttcccccagc aagctcgcgt cgtgcaggac gtgacaaatg gaagtagcac
     2521 gtctcactag tctcgtgcag atggacagca ccgctgagca atggaagcgg gtaggccttt
     2581 ggggcagcgg ccaatagcag ctttgttcct tcgctttctg ggctcagagg ctgggaaggg
     2641 gtgggtccgg gggcgggctc aggggcgggc tcaggggcgg gcgggcgccc gaaggtcctc
     2701 ccgaggcccg gcattctgca cgcttcaaaa gcgcacgtct gccgcgctgt tctcctcttc
     2761 ctcatctccg ggcctttcga cctgcagctc gaggtcgacc atgcccaaga agaagaggaa
     2821 ggtgtccaat ttactgaccg tacaccaaaa tttgcctgca ttaccggtcg atgcaacgag
     2881 tgatgaggtt cgcaagaacc tgatggacat gttcagggat cgccaggcgt tttctgagca
     2941 tacctggaaa atgcttctgt ccgtttgccg gtcgtgggcg gcatggtgca agttgaataa
     3001 ccggaaatgg tttcccgcag aacctgaaga tgttcgcgat tatcttctat atcttcaggc
     3061 gcgcggtctg gcagtaaaaa ctatccagca acatttgggc cagctaaaca tgcttcatcg
     3121 tcggtccggg ctgccacgac caagtgacag caatgctgtt tcactggtta tgcggcggat
     3181 ccgaaaagaa aacgttgatg ccggtgaacg tgcaaaacag gctctagcgt tcgaacgcac
     3241 tgatttcgac caggttcgtt cactcatgga aaatagcgat cgctgccagg atatacgtaa
     3301 tctggcattt ctggggattg cttataacac cctgttacgt atagccgaaa ttgccaggat
     3361 cagggttaaa gatatctcac gtactgacgg tgggagaatg ttaatccata ttggcagaac
     3421 gaaaacgctg gttagcaccg caggtgtaga gaaggcactt agcctggggg taactaaact
     3481 ggtcgagcga tggatttccg tctctggtgt agctgatgat ccgaataact acctgttttg
     3541 ccgggtcaga aaaaatggtg ttgccgcgcc atctgccacc agccagctat caactcgcgc
     3601 cctggaaggg atttttgaag caactcatcg attgatttac ggcgctaagg atgactctgg
     3661 tcagagatac ctggcctggt ctggacacag tgcccgtgtc ggagccgcgc gagatatggc
     3721 ccgcgctgga gtttcaatac cggagatcat gcaagctggt ggctggacca atgtaaatat
     3781 tgtcatgaac tatatccgta acctggatag tgaaacaggg gcaatggtgc gcctgctgga
     3841 agatggcgat tagccattaa cgcgtaaatg attgcagatc cactagttct agagctcgct
     3901 gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc
     3961 cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg
     4021 catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca
     4081 agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatggctt
     4141 ctgaggaaag aaccagctgg ggctcgagat ccactagttc tagcctcgag gctagagcgg
     4201 ccgccaccgc ggtggagctc caattcgccc tatagtgagt cgtattacgc gcgctcactg
     4261 gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg ttacccaact taatcgcctt
     4321 gcagcacatc cccctttcgc cagctggcgt aatagcgaag aggcccgcac cgatcgccct
     4381 tcccaacagt tgcgcagcct gaatggcgaa tgggacgcgc cctgtagcgg cgcattaagc
     4441 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc
     4501 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
     4561 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
     4621 aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
     4681 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
     4741 ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat
     4801 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg
     4861 cttacaattt
//
