LOCUS       sequence-254371-        4685 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian, Zebrafish, or Xenopus expression of PIF6CreN, one half of
            the CreLite system.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4685)
  AUTHORS   Yen ST, Trimmer KA, Aboul-Fettouh N, Mullen RD, Culver JC, Dickinson
            ME, Behringer RR, Eisenhoffer GT
  TITLE     CreLite: An optogenetically controlled Cre/loxP system using red 
            light.
  JOURNAL   Dev Dyn. 2020 Nov;249(11):1394-1403. doi: 10.1002/dvdy.232. Epub 
            2020 Aug 31.
  PUBMED    32745301
REFERENCE   2  (bases 1 to 4685)
  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: "10.1002/dvdy.232"; 
            journalName: "Dev Dyn"; date: "2020-11"; volume: "249"; issue: "11";
            pages: "1394-1403"
COMMENT     Sequence Label: pCS2+PIF6CreN
FEATURES             Location/Qualifiers
     source          1..4685
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        35..53
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     35..52
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             103..123
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    734..868
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(758..777)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     812..831
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(981..1001)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(981..999)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1020..1036)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1020..1036)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1033..1055)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1044..1060
                     /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)."
     promoter        complement(join(1068..1074,1075..1092,1093..1098))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1113..1134
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1251..1268)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1422..2010)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1502..1521)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2181..2972,2973..3041))
                     /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     2804..2823
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3042..3146)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3172..3627)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3309..3330)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3521..3540
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3701..4685
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
ORIGIN
        1 cgccattctg cctggggacg tcggagcaag cttgatttag gtgacactat agaatacaag
       61 ctacttgttc tttttgcagg atcccatcga ttcgaattca tgcccaagaa gaagaggaag
      121 gtgatgttct taccaaccga ttattgttgc aggttaagcg atcaagagta tatggagctt
      181 gtgtttgaga atggccagat tcttgcaaag ggccaaagat ccaacgtttc tctgcataat
      241 caacgtacca aatcgatcat ggatttgtat gaggcagagt ataacgagga tttcatgaag
      301 agtatcatcc atggtggtgg tggtgccatc acaaatctcg gggacacgca ggttgttcca
      361 caaagtcatg ttgctgctgc ccatgaaaca aacatgttgg aaagcaataa acatgttgac
      421 ggtggttctg ccggtggctc cggttctggc tccagcggtg gcagctctgg tgcgtccggc
      481 acgggtactg cgggtggcac tggcagcggt tccggtactg gctctggcct gactgtgcac
      541 caaaacctgc ctgccctccc tgtggatgcc acctctgatg aagtcaggaa gaacctgatg
      601 gacatgttca gggacaggca ggccttctct gaacacacct ggaagatgct cctgtctgtg
      661 tgcagatcct gggctgcctg gtgcaagctg aactgactcg agcctctaga actatagtga
      721 gtcgtattac gtagatccag acatgataag atacattgat gagtttggac aaaccacaac
      781 tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt
      841 aaccattata agctgcaata aacaagttaa caacaacaat tgcattcatt ttatgtttca
      901 ggttcagggg gaggtgtggg aggtttttta attcgcggcc gcggcgccaa tgcattgggc
      961 ccggtaccca gcttttgttc cctttagtga gggttaattg cgcgcttggc gtaatcatgg
     1021 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
     1081 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
     1141 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     1201 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     1261 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     1321 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     1381 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     1441 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     1501 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     1561 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     1621 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     1681 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     1741 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     1801 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     1861 aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     1921 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     1981 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     2041 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     2101 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     2161 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     2221 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     2281 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     2341 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     2401 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     2461 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     2521 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     2581 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     2641 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     2701 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     2761 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     2821 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     2881 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     2941 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     3001 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     3061 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     3121 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc taaattgtaa
     3181 gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc
     3241 aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga
     3301 gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag
     3361 ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt
     3421 ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta
     3481 gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag
     3541 cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg
     3601 cgcttaatgc gccgctacag ggcgcgtccc attcgccatt caggctgcgc aactgttggg
     3661 aagggcgatc ggtgcgggcc tcttcgctat tacgccagtc gaccatagcc aattcaatat
     3721 ggcgtatatg gactcatgcc aattcaatat ggtggatctg gacctgtgcc aattcaatat
     3781 ggcgtatatg gactcgtgcc aattcaatat ggtggatctg gaccccagcc aattcaatat
     3841 ggcggacttg gcaccatgcc aattcaatat ggcggacttg gcactgtgcc aactggggag
     3901 gggtctactt ggcacggtgc caagtttgag gaggggtctt ggccctgtgc caagtccgcc
     3961 atattgaatt ggcatggtgc caataatggc ggccatattg gctatatgcc aggatcaata
     4021 tataggcaat atccaatatg gccctatgcc aatatggcta ttggccaggt tcaatactat
     4081 gtattggccc tatgccatat agtattccat atatgggttt tcctattgac gtagatagcc
     4141 cctcccaatg ggcggtccca tataccatat atggggcttc ctaataccgc ccatagccac
     4201 tcccccattg acgtcaatgg tctctatata tggtctttcc tattgacgtc atatgggcgg
     4261 tcctattgac gtatatggcg cctcccccat tgacgtcaat tacggtaaat ggcccgcctg
     4321 gctcaatgcc cattgacgtc aataggacca cccaccattg acgtcaatgg gatggctcat
     4381 tgcccattca tatccgttct cacgccccct attgacgtca atgacggtaa atggcccact
     4441 tggcagtaca tcaatatcta ttaatagtaa cttggcaagt acattactat tggaaggacg
     4501 ccagggtaca ttggcagtac tcccattgac gtcaatggcg gtaaatggcc cgcgatggct
     4561 gccaagtaca tccccattga cgtcaatggg gaggggcaat gacgcaaatg ggcgttccat
     4621 tgacgtaaat gggcggtagg cgtgcctaat gggaggtcta tataagcaat gctcgtttag
     4681 ggaac
//
