LOCUS       sequence-10577-e        4280 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 4280)
  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 4280)
  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-EGFP
FEATURES             Location/Qualifiers
     source          1..4280
                     /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     59..79
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        61..79
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     regulatory      480..489
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(486..488,489..491,492..1205)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(531..552)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(792..811)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1139..1160
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    1247..1381
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1271..1290)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1325..1344
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1445..1461)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1446..1462)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     promoter        complement(1487..1505)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1533..1550)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1533..1549)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1542..1564)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      1691..2146
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1778..1797)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1988..2009
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2171..2275
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2276..2344,2345..3136)
                     /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(2494..2513)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3307..3895
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3796..3815
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4049..4066
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4183..4204
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4219..4224,4225..4242,4243..4249)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4257..4273
                     /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(4262..4280,1..4)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
ORIGIN
        1 caggaaacag ctatgaccat gattacgcca agcgcgtagg gataacaggg taatgcgcgc
       61 aattaaccct cactaaaggg aacaaaagct ggagctccac cgcggtggcg gccgctctag
      121 aactagtccg aagcttgcaa tgcctgcagg tcgactctag aggatgggct cgacggccca
      181 agctacatca agtgaccttt ttaaccaggt cagtgatttg tcatccattc gaaagctctg
      241 gagagtctaa gataaaacat acaataaccc acacctcccc catagtcatt atatagaaaa
      301 gagaggggaa aaaaagtgcc ctgtcccact gcgggcccct tttgtgctgt tcctgccaac
      361 acagcagacc tcctgctata tagaccctgc tcccagcccc acacactcaa cagcaccatc
      421 ccatccgacc tgccaacacc agccaagctt cccatcgatt cgaattcaag gagcttgccg
      481 ccaccatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc
      541 tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca
      601 cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc gtgccctggc
      661 ccaccctcgt gaccaccttc acctacggcg tgcagtgctt cagccgctac cccgaccaca
      721 tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca
      781 tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca
      841 ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg
      901 ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcatggcc gacaagcaga
      961 agaacggcat caaggtgaac ttcaagatcc gccacaacat cgaggacggc agcgtgcagc
     1021 tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca
     1081 accactacct gagcacccag tccgccctga gcaaagaccc caacgagaag cgcgatcaca
     1141 tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca
     1201 agtaaagcgg ccgcgactct agaactatag tgagtcgtat tacgtagatc cagacatgat
     1261 aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat
     1321 ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt
     1381 taacaacaac aattgcattc attttatgtt tcaggttcag ggggaggtgt gatatcaagc
     1441 ttatcgatac cgtcgacctc gagggggggc ccggtaccca attcgcccta tagtgagtcg
     1501 tattacgcgc gcattaccct gttatcccta tcactggccg tcgttttaca acgtcgtgac
     1561 tgggaaaacc ctggcgttac ccaacttaat cgccttgcag cacatccccc tttcgccagc
     1621 tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat
     1681 ggcgaatggg acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     1741 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc
     1801 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
     1861 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca
     1921 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
     1981 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct
     2041 tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa
     2101 caaaaattta acgcgaattt taacaaaata ttaacgctta caatttagtg gcacttttcg
     2161 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     2221 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
     2281 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     2341 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     2401 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     2461 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
     2521 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     2581 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     2641 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     2701 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     2761 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
     2821 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     2881 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     2941 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     3001 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     3061 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     3121 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     3181 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     3241 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     3301 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     3361 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     3421 tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca
     3481 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     3541 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     3601 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     3661 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     3721 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     3781 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     3841 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     3901 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     3961 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac
     4021 cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg
     4081 cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga
     4141 caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac
     4201 tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt
     4261 gagcggataa caatttcaca
//
