LOCUS       sequence-176065-        7137 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 7137)
  AUTHORS   Dor Y, Brown J, Martinez OI, Melton DA
  TITLE     Adult pancreatic beta-cells are formed by self-duplication rather 
            than stem-cell differentiation.
  JOURNAL   Nature. 2004 May 6. 429(6987):41-6.
  PUBMED    15129273
REFERENCE   2  (bases 1 to 7137)
  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: "Nature. 
            2004 May 6. 429(6987):41-6."
COMMENT     Sequence Label: RIP Cre-ER (DM#265)
FEATURES             Location/Qualifiers
     source          1..7137
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     85..102
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    219..240
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(255..260,261..278,279..285)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    293..309
                     /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     298..320
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     317..333
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     317..333
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     352..372
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        354..372
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     CDS             2064..3092
                     /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
                     LGIAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALSLGVTKLVE
                     RWISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYGAKDDSGQ
                     RYLAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRNLDSETGAMVRLLE
                     DGD"
     polyA_signal    4473..4607
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4510..4529)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4564..4583
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4679..4698)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4680..4698)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4705..4722)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4705..4721)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4714..4736)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4863..5318
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4950..4969)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5160..5181
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5344..5448
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5449..5517,5518..6309)
                     /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(5667..5686)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6480..7068
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6969..6988
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
       61 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca
      121 atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg
      181 tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat
      241 taggcacccc aggctttaca ctttatgctc ccggctcgta tgttgtgtgg aattgtgagc
      301 ggataacaat ttcacacagg aaacagctat gaccatgatt acgccaagcg cgcaattaac
      361 cctcactaaa gggaacaaaa gctgggtacg gccggccgta ccgggccccc cctcgaggtc
      421 gggggatccc ccaaccactc caagtggagg ctgagaaagg ttttgtagct gggtagagta
      481 tgtactaaga gatggagaca gctggctctg agctctgaag caagcacctc ttatggagag
      541 ttgctgacct tcaggtgcaa atctaagata ctacaggaga atacaccatg gggcttcagc
      601 ccagttgact cccgagtggg ctatgggttt gtggaaggag agatagaaga gaagggacct
      661 ttcttcttga attctgcttt ccttctacct ctgagggtga gctggggtct cagctgaggt
      721 gaggacacag ctatcagtgg gaactgtgaa acaacagttc aagggacaaa gttactaggt
      781 cccccaacaa ctgcagcctc ctggggaatg atgtggaaaa atgctcagcc aaggacaaag
      841 aaggcctcac cctctctgag acaatgtccc ctgctgtgaa ctggttcatc aggccaccca
      901 ggagccccta ttaagactct aattacccta aggctaagta gaggtgttgt tgtccaatga
      961 gcactttctg cagacctagc accaggcaag tgtttggaaa ctgcagcttc agcccctctg
     1021 gccatctgct gatccaccct taatgggaca aacagcaaag tccaggggtc aggggggggg
     1081 tgctttggac tataaagcta gtggggattc agtaaccccc agccctaact ctagagtcga
     1141 cctgcaggca tgcaagcttg atatcgaatt cctgcagccc cgcgggtacc gagctccagg
     1201 aacatccaaa ctgagcagcc ggggtccccc ccacccccca ccccgcccct cccggcaact
     1261 ttgagcctgt gctgggacag agcctctagt tcctaaatta gtccatgagg tcagaggcag
     1321 cactgccatt gtaacgcgat tggagaggat cacgtcaccg gacacgcccc caggcatctc
     1381 cctgggtctc ctaaacttgg cggggagaag ttttagccct taagttttag cctttaaccc
     1441 ccatattcag aactgtgcga gttggcgaaa ccccacaaat cacaacaaac tgtacacaac
     1501 accgagctag aggtgatctt tcttgtccat tccacacagg ccttagtaat gcgtcgccat
     1561 agcaacagtg tcactagtag caccagcact tccccacacc ctccccctca ggaatccgta
     1621 ctctccagtg aaccccagaa acctctggag agttctggac aagggcggaa cccacaactc
     1681 cgattactca agggaggcgg ggaagctcca ccagacgcga aactgctgga agattcctgg
     1741 ccccaaggcc tcctccggct cgctgattgg cccagcggag agtgggcggg gccggtgaag
     1801 actccttaaa ggcgcagggc ggcgagcagg tcaccagacg ctgacagcta ctcagaacca
     1861 aatctggttc catccagaga caagcgaaga caagagaagc agagcgagcg gcgcgttccc
     1921 gatcctcggc caggaccagc cttccccaga gcatccctgc cgcggagcgc aaccttccca
     1981 ggagcatccc tgccgcggag cgcaactttc cccggagcat ccacgccgcg gagcgcagcc
     2041 ttccagaagc agagcgcggc gccatgtcca atttactgac cgtacaccaa aatttgcctg
     2101 cattaccggt cgatgcaacg agtgatgagg ttcgcaagaa cctgatggac atgttcaggg
     2161 atcgccaggc gttttctgag catacctgga aaatgcttct gtccgtttgc cggtcgtggg
     2221 cggcatggtg caagttgaat aaccggaaat ggtttcccgc agaacctgaa gatgttcgcg
     2281 attatcttct atatcttcag gcgcgcggtc tggcagtaaa aactatccag caacatttgg
     2341 gccagctaaa catgcttcat cgtcggtccg ggctgccacg accaagtgac agcaatgctg
     2401 tttcactggt tatgcggcgg atccgaaaag aaaacgttga tgccggtgaa cgtgcaaaac
     2461 aggctctagc gttcgaacgc actgatttcg accaggttcg ttcactcatg gaaaatagcg
     2521 atcgctgcca ggatatacgt aatctggcat ttctggggat tgcttataac accctgttac
     2581 gtatagccga aattgccagg atcagggtta aagatatctc acgtactgac ggtgggagaa
     2641 tgttaatcca tattggcaga acgaaaacgc tggttagcac cgcaggtgta gagaaggcac
     2701 ttagcctggg ggtaactaaa ctggtcgagc gatggatttc cgtctctggt gtagctgatg
     2761 atccgaataa ctacctgttt tgccgggtca gaaaaaatgg tgttgccgcg ccatctgcca
     2821 ccagccagct atcaactcgc gccctggaag ggatttttga agcaactcat cgattgattt
     2881 acggcgctaa ggatgactct ggtcagagat acctggcctg gtctggacac agtgcccgtg
     2941 tcggagccgc gcgagatatg gcccgcgctg gagtttcaat accggagatc atgcaagctg
     3001 gtggctggac caatgtaaat attgtcatga actatatccg taacctggat agtgaaacag
     3061 gggcaatggt gcgcctgctg gaagatgggg atcaggctgg tgccatgggc gatccacgaa
     3121 atgaaatggg tgcttcagga gacatgaggg ctgccaacct ttggccaagc cctcttgtga
     3181 ttaagcacac taagaagaat agccctgcct tgtccttgac agctgaccag atggtcagtg
     3241 ccttgttgga tgctgaaccg cccatgatct attctgaata tgatccttct agacccttca
     3301 gtgaagcctc aatgatgggc ttattgacca acctagcaga tagggagctg gttcatatga
     3361 tcaactgggc aaagagagtg ccaggctttg gggacttgaa tctccatgat caggtccacc
     3421 ttctcgagtg tgcctggctg gagattctga tgattggtct cgtctggcgc tccatggaac
     3481 acccggggaa gctcctgttt gctcctaact tgctcctgga caggaatcaa ggtaaatgtg
     3541 tggaaggcat ggtggagatc tttgacatgt tgcttgctac gtcaagtcgg ttccgcatga
     3601 tgaacctgca gggtgaagag tttgtgtgcc tcaaatccat cattttgctt aattccggag
     3661 tgtacacgtt tctgtccagc accttgaagt ctctggaaga gaaggaccac atccaccgtg
     3721 tcctggacaa gatcacagac actttgatcc acctgatggc caaagctggc ctgactctgc
     3781 agcagcagca tcgccgccta gctcagctcc ttctcattct ttcccatatc cggcatatga
     3841 gtaacaaacg catggagcat ctctacaaca tgaaatgcaa gaacgtggta cccctctatg
     3901 acctgctcct ggagatgttg gatgcccacc gccttcatgc cccagccagt cgcatgggag
     3961 tgcccccaga ggagcccagc cagacccagc tggccaccac cagctccact tcagcacatt
     4021 ccttacaaac ctactacata cccccggaag cagagggctt ccccaacacg atctgagagc
     4081 tcctactagt ctagcttggg atctttgtga aggaacctta cttctgtggt gtgacataat
     4141 tggacaaact acctacagag atttaaagct ctaaggtaaa tataaaattt ttaagtgtat
     4201 aatgtgttaa actagctgca tatgcttgct gcttgagagt tttgcttact gagtatgatt
     4261 tatgaaaata ttatacacag gagctagtga ttctaattgt ttgtgtattt tagattcaca
     4321 gtcccaaggc tcatttcagg cccctcagtc ctcacagtct gttcatgatc ataatcagcc
     4381 ataccacatt tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc
     4441 tgaaacataa aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt
     4501 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     4561 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctggatcgat ctgcggccgc
     4621 caccgcggtg gagctgttta aacagctgtt taaacagctg tttaaacgct ccaattcgcc
     4681 ctatagtgag tcgtattaca attcactggc cgtcgtttta caacgtcgtg actgggaaaa
     4741 ccctggcgtt acccaactta atcgccttgc agcacatccc cctttcgcca gctggcgtaa
     4801 tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg
     4861 ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     4921 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     4981 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     5041 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg
     5101 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     5161 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
     5221 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
     5281 taacgcgaat tttaacaaaa tattaacgct tacaatttag gtggcacttt tcggggaaat
     5341 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     5401 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa
     5461 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
     5521 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
     5581 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
     5641 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc
     5701 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
     5761 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
     5821 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
     5881 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
     5941 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg
     6001 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
     6061 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
     6121 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt
     6181 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
     6241 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
     6301 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat
     6361 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     6421 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     6481 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     6541 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     6601 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
     6661 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     6721 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     6781 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     6841 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     6901 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     6961 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     7021 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     7081 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcct
//
