LOCUS       sequence-10575-e        4244 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 4244)
  AUTHORS   Vopalensky P, Ruzickova J, Pavlu B, Kozmik Z
  TITLE     A lens-specific co-injection marker for medaka transgenesis.
  JOURNAL   Biotechniques. 2010 Mar . 48(3):235-6.
  PUBMED    20359307
REFERENCE   2  (bases 1 to 4244)
  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: 
            "Biotechniques. 2010 Mar . 48(3):235-6."
COMMENT     Sequence Label: p817-mgammaFcry-mCherry
FEATURES             Location/Qualifiers
     source          1..4244
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..17
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1..17
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     54..74
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        56..74
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     CDS             467..1177
                     /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(612..630)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     880..899
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    1211..1345
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1235..1254)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1289..1308
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1409..1425)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1410..1426)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     promoter        complement(1451..1469)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1497..1514)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1497..1513)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1506..1528)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      1655..2110
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1742..1761)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1952..1973
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2135..2239
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2240..2308,2309..3100)
                     /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(2458..2477)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3271..3859
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3760..3779
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4013..4030
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4147..4168
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4183..4188,4189..4206,4207..4213)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4221..4237
                     /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     join(4226..4244,1..4)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
ORIGIN
        1 caggaaacag ctatgaccat gattacgcca atagggataa cagggtaatg cgcgcaatta
       61 accctcacta aagggaacaa aagctggagc tccaccgcgg tggcggccgc tctagaacta
      121 gtccgaagct tgcaatgcct gcaggtcgac tctagaggat gggctcgacg gcccaagcta
      181 catcaagtga cctttttaac caggtcagtg atttgtcatc cattcgaaag ctctggagag
      241 tctaagataa aacatacaat aacccacacc tcccccatag tcattatata gaaaagagag
      301 gggaaaaaaa gtgccctgtc ccactgcggg ccccttttgt gctgttcctg ccaacacagc
      361 agacctcctg ctatatagac cctgctccca gccccacaca ctcaacagca ccatcccatc
      421 cgacctgcca acaccagcca agcttcccat cgattcgaat tccaccatgg tgagcaaggg
      481 cgaggaggat aacatggcca tcatcaagga gttcatgcgc ttcaaggtgc acatggaggg
      541 ctccgtgaac ggccacgagt tcgagatcga gggcgagggc gagggccgcc cctacgaggg
      601 cacccagacc gccaagctga aggtgaccaa gggtggcccc ctgcccttcg cctgggacat
      661 cctgtcccct cagttcatgt acggctccaa ggcctacgtg aagcaccccg ccgacatccc
      721 cgactacttg aagctgtcct tccccgaggg cttcaagtgg gagcgcgtga tgaacttcga
      781 ggacggcggc gtggtgaccg tgacccagga ctcctccctg caggacggcg agttcatcta
      841 caaggtgaag ctgcgcggca ccaacttccc ctccgacggc cccgtaatgc agaagaagac
      901 catgggctgg gaggcctcct ccgagcggat gtaccccgag gacggcgccc tgaagggcga
      961 gatcaagcag aggctgaagc tgaaggacgg cggccactac gacgctgagg tcaagaccac
     1021 ctacaaggcc aagaagcccg tgcagctgcc cggcgcctac aacgtcaaca tcaagttgga
     1081 catcacctcc cacaacgagg actacaccat cgtggaacag tacgaacgcg ccgagggccg
     1141 ccactccacc ggcggcatgg acgagctgta caagtaactc gactagaact atagtgagtc
     1201 gtattacgta gatccagaca tgataagata cattgatgag tttggacaaa ccacaactag
     1261 aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac
     1321 cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt
     1381 tcagggggag gtgtgatatc aagcttatcg ataccgtcga cctcgagggg gggcccggta
     1441 cccaattcgc cctatagtga gtcgtattac gcgcgcatta ccctgttatc cctatcactg
     1501 gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg ttacccaact taatcgcctt
     1561 gcagcacatc cccctttcgc cagctggcgt aatagcgaag aggcccgcac cgatcgccct
     1621 tcccaacagt tgcgcagcct gaatggcgaa tgggacgcgc cctgtagcgg cgcattaagc
     1681 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc
     1741 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
     1801 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
     1861 aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
     1921 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
     1981 ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat
     2041 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg
     2101 cttacaattt agtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
     2161 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     2221 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
     2281 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
     2341 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
     2401 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct
     2461 atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca
     2521 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
     2581 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
     2641 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
     2701 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
     2761 cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg
     2821 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
     2881 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
     2941 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
     3001 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     3061 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc
     3121 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     3181 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     3241 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     3301 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     3361 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgtcct
     3421 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     3481 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     3541 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     3601 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     3661 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     3721 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     3781 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     3841 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     3901 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
     3961 taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc
     4021 agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc
     4081 gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa
     4141 cgcaattaat gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc
     4201 ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt caca
//
