LOCUS       sequence-11366-e        7905 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7905)
  AUTHORS   Hesselson D, Anderson RM, Beinat M, Stainier DY
  TITLE     Distinct populations of quiescent and proliferative pancreatic 
            beta-cells identified by HOTcre mediated labeling.
  JOURNAL   Proc Natl Acad Sci U S A. 2009 Sep 1. 106(35):14896-901.
  PUBMED    19706417
REFERENCE   2  (bases 1 to 7905)
  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"; journalName: "Proc Natl 
            Acad Sci U S A. 2009 Sep 1. 106(35):14896-901."
COMMENT     Sequence Label: ins-cre_cryaa-Venus
FEATURES             Location/Qualifiers
     source          1..7905
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     43..62
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        43..61
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1253..1273
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1274..1981
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(1419..1437)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1687..1706
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             1994..3025
                     /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="MSNLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAFSEHTWKML
                     LSVCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNMLHRRSGL
                     PRPSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRCQDIRNLAF
                     LGIAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALSLGVTKLVX
                     RWISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYGAKDDSGQ
                     RYLAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRNLDSETGAMVRLLE
                     DGD"
     polyA_signal    3323..3457
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3360..3379)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3414..3433
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    3475..3522
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     polyA_signal    3546..3753
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     3742..3759
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(join(3777..4490,4491..4493,4494..4496))
                     /codon_start=1
                     /product="yellow fluorescent protein (YFP) with fast and 
                     efficient maturation (Nagai et al., 2002)"
                     /label=Venus
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3822..3843)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     4171..4190
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4430..4451
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5086..5106)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(5086..5104)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5137..5154)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5137..5153)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5146..5168)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5295..5750
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5382..5401)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5592..5613
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5776..5880
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5881..5949,5950..6741)
                     /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(6099..6118)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6912..7500
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7401..7420
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7654..7671
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7788..7809
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7824..7829,7830..7847,7848..7854)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7862..7878
                     /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     7867..7889
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7886..7902
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7886..7902
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 gattacgcca agcgcgttag ggataacagg gtaatgcgcg cgtaatacga ctcactatag
       61 ggcgaattgg agctcacgcg taagctttgt ttgtaccata ataagctcat tatataatat
      121 ttaaaatgat actgcatcat taattgatat gttttctagt tatttttaaa tattagcaac
      181 taaattagga aattatccat agtaaattta atgtaatatt atttgttata tttaacttca
      241 gcccacagtc tagtttagtt gttggtccct tatgagaaaa aatttgggca cctctgctct
      301 agtgcttgta aacatttttg agtatctata gttgaacatg aaagcatttt tttttttttt
      361 tttttaagaa aatgaagaaa gctaaaaacc tgtatccact gacttccatt aaaaaaagca
      421 aatactaggg atgtctatgg tgtcttcttt tgtgttcaac aaataagcag aaagtaactg
      481 ctgacagaac attacatttt gtgtgaatcc ccttatattg tgttaaaaac tgtagagaaa
      541 tcatctaata ataagaaatt atcaagaaat aaacagaaaa tgatctaata gtatactata
      601 aaatctttca aaatccaggc cactaccata tgactaaaac cacacatatt atgctttaaa
      661 aaaagtcaca atgaagacgt cacaaacgtt cctgaagact aatgactcgt ctaaccccat
      721 cttaccttct gctgtttcac tcaaagaaaa gccaccagct gtctctgtga tctgctcatt
      781 ctgtgttaac ttgtgttctc aacaattaac aattaaccaa aagcctctgg agctgcgtga
      841 gcatcacgat gtgtttgtct tctgtgagcg tggcagtgtc cacgaagtca atgtgaattg
      901 gttggccaaa cacagctaat gtggcatgtg tgattggaca gccacttaaa agtgttaaag
      961 tgtgtttacg cacaggattg gttttccgct gaatcaggca aaaccagcag aaccgatcta
     1021 aactcgagta taaaagtgag taggagatgc tattcgtccc aaaacatctc caccaccatc
     1081 tccaccattc ctcgcctctg cttcgagaac aggtgagtgt cgagcgggat ggtaaatcta
     1141 cagagaatgc ggagtgtagc ttgtgtacat gtttttgatt aacagagatt gtatgtgtgt
     1201 gtttgtgtca gtgtgacgga tctaattcga tatcaagctt atcgatatgg ctccaaagaa
     1261 gaagcgtaag gtaatggtga gcaagggcga ggaggataac atggccatca tcaaggagtt
     1321 catgcgcttc aaggtgcaca tggagggctc cgtgaacggc cacgagttcg agatcgaggg
     1381 cgagggcgag ggccgcccct acgagggcac ccagaccgcc aagctgaagg tgaccaaggg
     1441 tggccccctg cccttcgcct gggacatcct gtcccctcag ttcatgtacg gctccaaggc
     1501 ctacgtgaag caccccgccg acatccccga ctacttgaag ctgtccttcc ccgagggctt
     1561 caagtgggag cgcgtgatga acttcgagga cggcggcgtg gtgaccgtga cccaggactc
     1621 ctccctgcag gacggcgagt tcatctacaa ggtgaagctg cgcggcacca acttcccctc
     1681 cgacggcccc gtaatgcaga agaagaccat gggctgggag gcctcctccg agcggatgta
     1741 ccccgaggac ggcgccctga agggcgagat caagcagagg ctgaagctga aggacggcgg
     1801 ccactacgac gctgaggtca agaccaccta caaggccaag aagcccgtgc agctgcccgg
     1861 cgcctacaac gtcaacatca agttggacat cacctcccac aacgaggact acaccatcgt
     1921 ggaacagtac gaacgcgccg agggccgcca ctccaccggc ggcatggacg agctgtacaa
     1981 ggcagctagc gcaatgtcca atttactgac cgtacaccaa aatttgcctg cattaccggt
     2041 cgatgcaacg agtgatgagg ttcgcaagaa cctgatggac atgttcaggg atcgccaggc
     2101 gttttctgag catacctgga aaatgcttct gtccgtttgc cggtcgtggg cggcatggtg
     2161 caagttgaat aaccggaaat ggtttcccgc agaacctgaa gatgttcgcg attatcttct
     2221 atatcttcag gcgcgcggtc tggcagtaaa aactatccag caacatttgg gccagctaaa
     2281 catgcttcat cgtcggtccg ggctgccacg accaagtgac agcaatgctg tttcactggt
     2341 tatgcggcgg atccgaaaag aaaacgttga tgccggtgaa cgtgcaaaac aggctctagc
     2401 gttcgaacgc actgatttcg accaggttcg ttcactcatg gaaaatagcg atcgctgcca
     2461 ggatatacgt aatctggcat ttctggggat tgcttataac accctgttac gtatagccga
     2521 aattgccagg atcagggtta aagatatctc acgtactgac ggtgggagaa tgttaatcca
     2581 tattggcaga acgaaaacgc tggttagcac cgcaggtgta gagaaggcac ttagcctggg
     2641 ggtaactaaa ctggtcnagc gatggatttc cgtctctggt gtagctgatg atccgaataa
     2701 ctacctgttt tgccgggtca gaaaaaatgg tgttgccgcg ccatctgcca ccagccagct
     2761 atcaactcgc gccctggaag ggatttttga agcaactcat cgattgattt acggcgctaa
     2821 ggatgactct ggtcagagat acctggcctg gtctggacac agtgcccgtg tcggagccgc
     2881 gcgagatatg gcccgcgctg gagtttcaat accggagatc atgcaagctg gtggctggac
     2941 caatgtaaat attgtcatga actatatccg taacctggat agtgaaacag gggcaatggt
     3001 gcgcctgctg gaagatggcg attaggcggc cgccgaaatg accgaccaag cgacgcccaa
     3061 cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat
     3121 cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt
     3181 cgcccacccc ccggatctaa gctctagata agtaatgatc ataatcagcc atatcacatc
     3241 tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc tgaaacataa
     3301 aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt acaaataaag
     3361 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     3421 gtccaaactc atcaatgtat cttatcatgt ctggatccgg gggtaccgcg tcgagaagtt
     3481 cctattccga agttcctatt ctctagaaag tataggaact tcgggcccgc ggtggcggcg
     3541 atctctcccc agcatgcctg ctattgtctt cccaatcctc ccccttgctg tcctgcccca
     3601 ccccaccccc cagaatagaa tgacacctac tcagacaatg cgatgcaatt tcctcatttt
     3661 attaggaaag gacagtggga gtggcacctt ccagggtcaa ggaaggcacg ggggaggggc
     3721 aaacaacaga tggctggcaa ctagaaggca cagtcgaggc tgatcagcgg gccgctttac
     3781 ttgtacagct cgtccatgcc gagagtgatc ccggcggcgg tcacgaactc cagcaggacc
     3841 atgtgatcgc gcttctcgtt ggggtctttg ctcagggcgg actggtagct caggtagtgg
     3901 ttgtcgggca gcagcacggg gccgtcgccg atgggggtgt tctgctggta gtggtcggcg
     3961 agctgcacgc cgccgtcctc gatgttgtgg cggatcttga agttggcctt gatgccgttc
     4021 ttctgcttgt cggcggtgat atagacgttg tggctgttgt agttgtactc cagcttgtgc
     4081 cccaggatgt tgccgtcctc cttgaagtcg atgcccttca gctcgatgcg gttcaccagg
     4141 gtgtcgccct cgaacttcac ctcggcgcgg gtcttgtagt tgccgtcgtc cttgaagaag
     4201 atggtgcgct cctggacgta gccttcgggc atggcggact tgaagaagtc gtgctgcttc
     4261 atgtggtcgg ggtagcgggc gaagcactgc aggccgtagc ccagggtggt cacgagggtg
     4321 ggccagggca cgggcagctt gccggtggtg cagatcagct tcagggtcag cttgccgtag
     4381 gtggcatcgc cctcgccctc gccggacacg ctgaacttgt ggccgtttac gtcgccgtcc
     4441 agctcgacca ggatgggcac caccccggtg aacagctcct cgcccttgct caccatgttg
     4501 tcagacctgg taactcctta ctgtaactta ttgcacctga tgaacaaaag atgtgaactg
     4561 aacgcaagta ggaggctaat agaggctagt acagtacaaa gatccccaaa tgatgtgtgc
     4621 cggccttagt ggaggcagac ccctggttat atacctgtgc tgcagaaaga ggggcttaca
     4681 aatggatctc tctggaatgg cattatctca tggtgggaac agtgttgtca gcagaatcag
     4741 cggggtggac tctgacctga gaatgacagt ctagcactga atggctcaga caatactctt
     4801 gtggccaact atcaccggaa aataagtgcc tgccaaaaca aaccagacca gatgaagcta
     4861 catggctgta gaacgtcagt atttaatgtg ttgtggccac ttggttcacc agatttaaga
     4921 gctaattttt ttaaatatca cagataatca atcgcttttt ttttttcttg actgaaaagc
     4981 ttagactgga aaattctttg aaaaagtttg cactacagaa aataaataca taacatcctg
     5041 cactgaatgc acactattaa tgggcccggt acccagcttt tgttcccttt agtgagggtt
     5101 aattgcgcgc tagggataac agggtaatac gcgctcactg gccgtcgttt tacaacgtcg
     5161 tgactgggaa aaccctggcg ttacccaact taatcgcctt gcagcacatc cccctttcgc
     5221 cagctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct
     5281 gaatggcgaa tgggacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
     5341 gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc
     5401 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt
     5461 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg
     5521 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
     5581 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta
     5641 ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat
     5701 ttaacaaaaa tttaacgcga attttaacaa aatattaacg cttacaattt aggtggcact
     5761 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     5821 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
     5881 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct
     5941 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca
     6001 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc
     6061 gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc
     6121 cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg
     6181 gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta
     6241 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc
     6301 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt
     6361 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg
     6421 cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct
     6481 tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc
     6541 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct
     6601 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac
     6661 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc
     6721 tcactgatta agcattggta actgtcagac caagtttact catatatact ttagattgat
     6781 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
     6841 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     6901 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     6961 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     7021 gtaactggct tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta
     7081 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     7141 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     7201 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     7261 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     7321 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     7381 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     7441 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     7501 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     7561 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
     7621 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
     7681 gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg
     7741 cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag
     7801 ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga
     7861 attgtgagcg gataacaatt tcacacagga aacagctatg accat
//
