LOCUS       sequence-57104-e        4880 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 4880)
  TITLE     inducible Drosophila S2 vector
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4880)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pCGMbX
FEATURES             Location/Qualifiers
     source          1..4880
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     polyA_signal    240..374
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(join(400..1113,1114..1116,1117..1119))
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             complement(join(403..1113,1114..1116,1117..1119))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(445..466)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     794..813
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1053..1074
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     1608..1630
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1623..1639
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        1645..2071
                     /label=MT promoter
                     /note="Drosophila metallothionein promoter"
     sig_peptide     2089..2142
                     /codon_start=1
                     /product="Drosophila BiP signal sequence"
                     /label=BiP signal sequence
                     /translation="MKLCILLAVVAFVGLSLG"
     CDS             2241..2282
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             2292..2309
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2332..2349)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2382..2516
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2660..2676)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2660..2676)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2673..2695)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2684..2700
                     /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(2708..2714,2715..2732,2733..2738))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2753..2774
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2891..2908)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3062..3650)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3142..3161)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3821..4612,4613..4681))
                     /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     4444..4463
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4682..4786)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4854..4872
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcccg
      241 atccagacat gataagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa
      301 aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct
      361 gcaataaaca agttcaagct agctggcctc gaggccgctt tacttgtaca gctcgtccat
      421 gccgagagtg atcccggcgg cggtcacgaa ctccagcagg accatgtgat cgcgcttctc
      481 gttggggtct ttgctcaggg cggactgggt gctcaggtag tggttgtcgg gcagcagcac
      541 ggggccgtcg ccgatggggg tgttctgctg gtagtggtcg gcgagctgca cgctgccgtc
      601 ctcgatgttg tggcggatct tgaagttcac cttgatgccg ttcttctgct tgtcggccat
      661 gatatagacg ttgtggctgt tgtagttgta ctccagcttg tgccccagga tgttgccgtc
      721 ctccttgaag tcgatgccct tcagctcgat gcggttcacc agggtgtcgc cctcgaactt
      781 cacctcggcg cgggtcttgt agttgccgtc gtccttgaag aagatggtgc gctcctggac
      841 gtagccttcg ggcatggcgg acttgaagaa gtcgtgctgc ttcatgtggt cggggtagcg
      901 gctgaagcac tgcacgccgt aggtcagggt ggtcacgagg gtgggccagg gcacgggcag
      961 cttgccggtg gtgcagatga acttcagggt cagcttgccg taggtggcat cgccctcgcc
     1021 ctcgccggac acgctgaact tgtggccgtt tacgtcgccg tccagctcga ccaggatggg
     1081 caccaccccg gtgaacagct cctcgccctt gctcaccatg gggggatctg ttgtaattta
     1141 taatttatat ttcctttctt aataaataaa taaatagtca agtttatgtt tgagttttat
     1201 gatttatatt ttaagttatt tcaactgcaa caccagcacc acgacctact tacagcaaaa
     1261 aacgtacaag aaggaaagaa ggaataaaaa gagtggtatt ctcttacaat atgttttatg
     1321 gcataaaagg tgtggccatt catatcaaat ataaagtagt gttgtttaac gttacttttg
     1381 taggttgaat agtatattcc aacagatgat gaggggttcc caatcctaaa cccatttgcc
     1441 gttcccagaa gcatgaaacc accacgcacc ggatcctcta ggccattcgc cattcaggct
     1501 gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa
     1561 agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg
     1621 ttgtaaaacg acggccagtg aattaattcg ttgcaggaca ggatgtggtg cccgatgtga
     1681 ctagctcttt gctgcaggcc gtcctatcct ctggttccga taagagaccc agaactccgg
     1741 ccccccaccg cccaccgcca cccccataca tatgtggtac gcaagtaaga gtgcctgcgc
     1801 atgccccatg tgccccacca agagctttgc atcccataca agtccccaaa gtggagaacc
     1861 gaaccaattc ttcgcgggca gaacaaaagc ttctgcacac gtctccactc gaatttggag
     1921 ccggccggcg tgtgcaaaag aggtgaatcg aacgaaagac ccgtgtgtaa agccgcgttt
     1981 ccaaaatgta taaaaccgag agcatctggc caatgtgcat cagttgtggt cagcagcaaa
     2041 atcaagtgaa tcatctcagt gcaactaaag gggggatccg atctcaatat gaagttatgc
     2101 atattactgg ccgtcgtggc ctttgttggc ctctcgctcg ggagatctcc atggcccggg
     2161 gtacctacta gtccagtgtg gtggaattct gcagatatcc agcacagtgg cggccgctcg
     2221 agtctagagg gcccttcgaa ggtaagccta tccctaaccc tctcctcggt ctcgattcta
     2281 cgcgtaccgg tcatcatcac catcaccatt gagtttaaac ccgctgatca gcctcgactg
     2341 tgccttctaa ggcctgagct cgctgatcag cctcgatcga ggatccagac atgataagat
     2401 acattgatga gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg
     2461 aaatttgtga tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca
     2521 acaacaattg cattcatttt atgtttcagg ttcaggggga ggtgtgggag gttttttaaa
     2581 gcaagtaaaa cctctacaaa tgtggtatgg ctgattatga tcagtcgacc tgcaggcatg
     2641 caagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
     2701 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
     2761 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     2821 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     2881 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     2941 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     3001 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     3061 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     3121 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     3181 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     3241 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     3301 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     3361 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     3421 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     3481 ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta
     3541 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     3601 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     3661 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     3721 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     3781 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     3841 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     3901 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     3961 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     4021 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     4081 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     4141 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     4201 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     4261 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     4321 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     4381 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     4441 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     4501 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     4561 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     4621 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     4681 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     4741 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     4801 aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc
     4861 gtatcacgag gccctttcgt
//
