LOCUS       sequence-241972-        4341 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 4341)
  TITLE     Other Plasmids
REFERENCE   2  (bases 1 to 4341)
  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"
COMMENT     Sequence Label: M38 TOP-dGFP
FEATURES             Location/Qualifiers
     source          1..4341
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    1..135
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        333..351
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     333..350
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     regulatory      436..445
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(442..444,445..447,448..1158,1159..1164,1165..1287)
                     /codon_start=1
                     /product="EGFP destabilized by fusion to residues 422-461 
                     of mouse ornithine decarboxylase, giving an in vivo 
                     half-life of ~2 hours"
                     /label=d2EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYKKLSHGFPPEVEEQDDGTLPMSCAQESGMDRHPAACASARINV
                     "
     primer_bind     complement(487..508)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(748..767)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1095..1116
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1319..1338)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1320..1338)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    1343..1477
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1590..1610)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1590..1608)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1629..1645)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1629..1645)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1642..1664)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1653..1669
                     /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(1677..1683,1684..1701,1702..1707))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1722..1743
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1860..1877)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2031..2619)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2111..2130)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2790..3581,3582..3650))
                     /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     3413..3432
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3651..3755)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3781..4236)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3918..3939)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4130..4149
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
       61 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
      121 tatcatgtct ggatcctcta gagtcgacct gcagcccaag ctatcaaagg gggtaagatc
      181 aaagggggta agatcaaagg gggtaagatc aaaggaggag cttcagtcga ccccagtgac
      241 gtaggaagtc catccattca cagcgcttct ataaaggcgc cagctgaggc gcctactact
      301 ccaaccgcga ctgcagcgag caactaagct tgatttaggt gacactatag aatacaagct
      361 acttgttctt tttgcaggat cccatcgatt cgaattctgc agtcgacggt accgcgggcc
      421 cgggatccac cggtcgccac catggtgagc aagggcgagg agctgttcac cggggtggtg
      481 cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag
      541 ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag
      601 ctgcccgtgc cctggcccac cctcgtgacc accctgacct acggcgtgca gtgcttcagc
      661 cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac
      721 gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg
      781 aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag
      841 gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc
      901 atggccgaca agcagaagaa cggcatcaag gtgaacttca agatccgcca caacatcgag
      961 gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc
     1021 gtgctgctgc ccgacaacca ctacctgagc acccagtccg ccctgagcaa agaccccaac
     1081 gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc
     1141 atggacgagc tgtacaagaa gcttagccat ggcttcccgc cggaggtgga ggagcaggat
     1201 gatggcacgc tgcccatgtc ttgtgcccag gagagcggga tggaccgtca ccctgcagcc
     1261 tgtgcttctg ctaggatcaa tgtgtaggcg gcccctctcg agcctctaga ttctgcagcc
     1321 ctatagtgag tcgtattacg tagatccaga catgataaga tacattgatg agtttggaca
     1381 aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc
     1441 tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt
     1501 tatgtttcag gttcaggggg aggtgtggga ggttttttaa ttcgcggccg cggcgccaat
     1561 gcattgggcc cggtacccag cttttgttcc ctttagtgag ggttaattgc gcgcttggcg
     1621 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     1681 atacgagccg ggagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
     1741 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     1801 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     1861 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     1921 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     1981 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     2041 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     2101 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     2161 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     2221 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     2281 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     2341 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     2401 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     2461 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     2521 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     2581 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     2641 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     2701 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     2761 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     2821 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     2881 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     2941 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     3001 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     3061 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     3121 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     3181 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     3241 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     3301 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     3361 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     3421 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     3481 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     3541 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     3601 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     3661 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     3721 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     3781 aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat
     3841 tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga
     3901 tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca
     3961 acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct
     4021 aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc
     4081 cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag
     4141 cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca
     4201 cacccgccgc gcttaatgcg ccgctacagg gcgcgtccca ttcgccattc aggctgcgca
     4261 actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagtcg agtgaaacat
     4321 aaatggaatg caattgttgt t
//
