LOCUS       sequence-308160-        5486 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 5486)
  AUTHORS   Nowotschin S, Hadjantonakis AK
  TITLE     Use of KikGR a photoconvertible green-to-red fluorescent protein for
            cell labeling and lineage analysis in ES cells and mouse embryos.
  JOURNAL   BMC Dev Biol. 2009 Sep 9;9:49.
  PUBMED    19740427
REFERENCE   2  (bases 1 to 5486)
  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: "BMC Dev 
            Biol."; date: "2009-09-9"; volume: "9"; pages: "49"
COMMENT     Sequence Label: pCAG:KikGR
FEATURES             Location/Qualifiers
     source          1..5486
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        382..656
                     /label=chicken beta-actin promoter
     intron          657..1673
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1597..1619
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1681..1700
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             1731..2417
                     /codon_start=1
                     /product="photoconvertible Kikume green-to-red fluorescent 
                     protein (Tsutsui et al., 2005)"
                     /label=KikGR1
                     /translation="MVSVITSEMKIELRMEGAVNGHKFVITGKGSGQPFEGIQNVDLTV
                     IEGGPLPFAFDILTTAFHYGNRVFVEYPEEIVDYFKQSFPEGYSWERSMSYEDGGICLA
                     TNNITMKKDGSNCFVNEIRFDGVNFPANGPVMQRKTVKWEPSTEKMYVRDGVLKGDVNM
                     ALLLQGGGHYRCDFRTTYKAKKVVQLPDYHFVDHQMEITSHDKDYNKVKLYEHAKAHSG
                     LPRLAK"
     primer_bind     complement(2504..2523)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2569..2624
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2623..2642)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(2985..3001)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2985..3001)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2998..3020)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3009..3025
                     /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(3033..3039,3040..3057,3058..3063))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3078..3099
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        3158..3353
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3204..3339
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    3359..3493
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3396..3415)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3450..3469
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3561..3578)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3732..4320)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3812..3831)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4491..5282,5283..5351))
                     /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     5114..5133
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5352..5456)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtcgaggtga gccccacgtt ctgcttcact ctccccatct
      421 cccccccctc cccaccccca attttgtatt tatttatttt ttaattattt tgtgcagcga
      481 tgggggcggg gggggggggg cgcgcgccag gcggggcggg gcggggcgag gggcggggcg
      541 gggcgaggcg gagaggtgcg gcggcagcca atcagagcgg cgcgctccga aagtttcctt
      601 ttatggcgag gcggcggcgg cggcggccct ataaaaagcg aagcgcgcgg cgggcgggag
      661 tcgctgcgcg ctgccttcgc cccgtgcccc gctccgccgc cgcctcgcgc cgcccgcccc
      721 ggctctgact gaccgcgtta ctcccacagg tgagcgggcg ggacggccct tctcctccgg
      781 gctgtaatta gcgcttggtt taatgacggc ttgtttcttt tctgtggctg cgtgaaagcc
      841 ttgaggggct ccgggagggc cctttgtgcg gggggagcgg ctcggggggt gcgtgcgtgt
      901 gtgtgtgcgt ggggagcgcc gcgtgcggct ccgcgctgcc cggcggctgt gagcgctgcg
      961 ggcgcggcgc ggggctttgt gcgctccgca gtgtgcgcga ggggagcgcg gccgggggcg
     1021 gtgccccgcg gtgcgggggg ggctgcgagg ggaacaaagg ctgcgtgcgg ggtgtgtgcg
     1081 tgggggggtg agcagggggt gtgggcgcgt cggtcgggct gcaacccccc ctgcaccccc
     1141 ctccccgagt tgctgagcac ggcccggctt cgggtgcggg gctccgtacg gggcgtggcg
     1201 cggggctcgc cgtgccgggc ggggggtggc ggcaggtggg ggtgccgggc ggggcggggc
     1261 cgcctcgggc cggggagggc tcgggggagg ggcgcggcgg cccccggagc gccggcggct
     1321 gtcgaggcgc ggcgagccgc agccattgcc ttttatggta atcgtgcgag agggcgcagg
     1381 gacttccttt gtcccaaatc tgtgcggagc cgaaatctgg gaggcgccgc cgcaccccct
     1441 ctagcgggcg cggggcgaag cggtgcggcg ccggcaggaa ggaaatgggc ggggagggcc
     1501 ttcgtgcgtc gccgcgccgc cgtccccttc tccctctcca gcctcggggc tgtccgcggg
     1561 gggacggctg ccttcggggg ggacggggca gggcggggtt cggcttctgg cgtgtgaccg
     1621 gcggctctag agcctctgct aaccatgttc atgccttctt ctttttccta cagctcctgg
     1681 gcaacgtgct ggttattgtg ctgtctcatc attttggcaa agaattccgg atggtgagtg
     1741 tgattacatc agaaatgaag atcgagctgc gtatggaagg cgctgtaaac gggcacaagt
     1801 tcgtgattac agggaaagga agtggccagc ctttcgaggg aatacagaat gtggacctga
     1861 cagtcataga gggcggacct cttccttttg ctttcgatat cctgacaaca gcattccatt
     1921 acggcaaccg ggtatttgtc gaatacccag aagaaatagt agactacttc aagcagtcgt
     1981 ttcctgaggg ttattcttgg gaacgaagca tgagttacga agacggggga atttgcctcg
     2041 ccacaaacaa tataacgatg aagaaagacg gcagcaactg ttttgtcaat gaaattcgat
     2101 ttgatggtgt gaactttcct gccaatggtc cagttatgca gaggaagacc gtcaaatggg
     2161 agccatccac tgagaaaatg tatgtgcgtg atggagtgct gaagggtgat gtaaacatgg
     2221 ctctgttgct tcaaggaggt ggccattacc gatgtgactt cagaactact tacaaagcaa
     2281 agaaggttgt ccagttgcca gactatcact tcgtggatca tcaaatggag ataacaagcc
     2341 atgacaagga ttacaacaag gttaagctgt atgagcacgc taaagctcat tccgggctgc
     2401 caaggctggc caagtaaccg gaattcactc ctcaggtgca ggctgcctat cagaaggtgg
     2461 tggctggtgt ggccaatgcc ctggctcaca aataccactg agatcttttt ccctctgcca
     2521 aaaattatgg ggacatcatg aagccccttg agcatctgac ttctggctaa taaaggaaat
     2581 ttattttcat tgcaatagtg tgttggaatt ttttgtgtct ctcactcgga aggacatatg
     2641 ggagggcaaa tcatttaaaa catcagaatg agtatttggt ttagagtttg gcaacatatg
     2701 cccatatgct ggctgccatg aacaaaggtt ggctataaag aggtcatcag tatatgaaac
     2761 agccccctgc tgtccattcc ttattccata gaaaagcctt gacttgaggt tagatttttt
     2821 ttatattttg ttttgtgtta tttttttctt taacatccct aaaattttcc ttacatgttt
     2881 tactagccag atttttcctc ctctcctgac tactcccagt catagctgtc cctcttctct
     2941 tatggagatc cctcgacctg cagcccaagc ttggcgtaat catggtcata gctgtttcct
     3001 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
     3061 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
     3121 gctttccagt cgggaaacct gtcgtgccag cggatccgca tctcaattag tcagcaacca
     3181 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
     3241 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg
     3301 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctaa
     3361 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     3421 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     3481 tcatgtctgg atccgctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     3541 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     3601 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     3661 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     3721 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     3781 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     3841 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     3901 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     3961 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     4021 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     4081 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     4141 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     4201 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     4261 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     4321 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     4381 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     4441 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     4501 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     4561 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     4621 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     4681 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     4741 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     4801 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     4861 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     4921 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     4981 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     5041 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     5101 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     5161 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     5221 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     5281 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     5341 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     5401 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     5461 tttccccgaa aagtgccacc tgggtc
//
