LOCUS       sequence-379513-        6634 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of BID-GFP chimera.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6634)
  AUTHORS   Scorrano L
  TITLE     Luca Scorrano plasmids
  JOURNAL   unpublished
REFERENCE   2  (bases 1 to 6634)
  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            "unpublished"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: pcDNA3.1-Bid-GFP
FEATURES             Location/Qualifiers
     source          1..6634
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1..795
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(55..74)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     665..684
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    969..1090
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1006..1025)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1060..1079
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1139..1155)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1139..1155)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1152..1174)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1163..1179
                     /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(1187..1193,1194..1211,1212..1217))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1232..1253
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1370..1387)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1541..2129)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1621..1640)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2300..3091,3092..3160))
                     /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     2923..2942
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3161..3265)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3340..3359)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        3531..3910
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3911..4114
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4065..4085
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     4159..4178
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4159..4177
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      4269..4278
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(4710..4712,4713..4715,4716..5429)
                     /codon_start=1
                     /product="Cycle 3 GFP (Crameri et al., 1996)"
                     /label=Cycle 3 GFP
                     /note="mammalian codon-optimized"
                     /translation="MASKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFSYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     primer_bind     complement(4743..4771)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     5366..5386
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     promoter        complement(5483..5501)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(5484..5501)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(5521..5538)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5527..5751
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      5797..6225
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5884..5903)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6094..6115
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6234..6254)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        6239..6568
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6419..6554
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6481..6500
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
ORIGIN
        1 atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc
       61 ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca
      121 gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg
      181 caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg
      241 ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag
      301 gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg
      361 cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc
      421 atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa
      481 gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac
      541 ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat
      601 ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac
      661 atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc
      721 ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt
      781 gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc
      841 tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg
      901 ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg
      961 cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     1021 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     1081 atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt
     1141 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
     1201 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
     1261 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     1321 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     1381 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     1441 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     1501 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     1561 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     1621 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     1681 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     1741 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     1801 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     1861 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     1921 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     1981 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     2041 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     2101 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     2161 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     2221 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     2281 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     2341 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     2401 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc
     2461 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     2521 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     2581 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     2641 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     2701 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     2761 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     2821 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     2881 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     2941 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     3001 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     3061 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     3121 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     3181 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     3241 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg
     3301 gatcgggaga tctcccgatc ccctatggtg cactctcagt acaatctgct ctgatgccgc
     3361 atagttaagc cagtatctgc tccctgcttg tgtgttggag gtcgctgagt agtgcgcgag
     3421 caaaatttaa gctacaacaa ggcaaggctt gaccgacaat tgcatgaaga atctgcttag
     3481 ggttaggcgt tttgcgctgc ttcgcgatgt acgggccaga tatacgcgtt gacattgatt
     3541 attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga
     3601 gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg
     3661 cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg
     3721 acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca
     3781 tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc
     3841 ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc
     3901 tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc
     3961 acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa
     4021 tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag
     4081 gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag aacccactgc
     4141 ttactggctt atcgaaatta atacgactca ctatagggag acccaagctt ggtaccgagc
     4201 tcggatccac tagtaacggc cgccagtgtg ctggaattct gcagatatcc atcacactgg
     4261 cggccgccgc caccatgggc agccaggcca gccgctcctt caaccaagga agaatagagc
     4321 cagattctga aagtcaggaa gaaatcatcc acaacattgc cagacatctc gcccaaatag
     4381 gcgatgagat ggaccacaac atccagccca cactggtgag acagctagcc gcacagttca
     4441 tgaatggcag cctgtcggag gaagacaaaa ggaactgcct ggccaaagcc cttgatgagg
     4501 tgaagacagc cttccccaga gacatggaga acgacaaggc catgctgata atgacaatgc
     4561 tgttggccaa aaaagtggcc agtcacgcac catctttgct ccgtgatgtc ttccacacga
     4621 ctgtcaactt tattaaccag aacctattct cctatgtgag gaacttggtt agaaacgaga
     4681 tggaccaagg gcaattctgc agatctagaa tggctagcaa aggagaagaa cttttcactg
     4741 gagttgtccc aattcttgtt gaattagatg gtgatgttaa tgggcacaaa ttttctgtca
     4801 gtggagaggg tgaaggtgat gctacatacg gaaagcttac ccttaaattt atttgcacta
     4861 ctggaaaact acctgttcca tggccaacac ttgtcactac tttctcttat ggtgttcaat
     4921 gcttttcccg ttatccggat catatgaaac ggcatgactt tttcaagagt gccatgcccg
     4981 aaggttatgt acaggaacgc actatatctt tcaaagatga cgggaactac aagacgcgtg
     5041 ctgaagtcaa gtttgaaggt gatacccttg ttaatcgtat cgagttaaaa ggtattgatt
     5101 ttaaagaaga tggaaacatt ctcggacaca aactcgagta caactataac tcacacaatg
     5161 tatacatcac ggcagacaaa caaaagaatg gaatcaaagc taacttcaaa attcgccaca
     5221 acattgaaga tggatccgtt caactagcag accattatca acaaaatact ccaattggcg
     5281 atggccctgt ccttttacca gacaaccatt acctgtcgac acaatctgcc ctttcgaaag
     5341 atcccaacga aaagcgtgac cacatggtcc ttcttgagtt tgtaactgct gctgggatta
     5401 cacatggcat ggatgagctc tacaaataat gaattatcca gcacagtggc ggccgctcga
     5461 gcatgcatct agagggccct attctatagt gtcacctaaa tgctagagct cgctgatcag
     5521 cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct
     5581 tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc
     5641 attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg
     5701 aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg
     5761 cggaaagaac cagctggggc tctagggggt atccccacgc gccctgtagc ggcgcattaa
     5821 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc
     5881 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag
     5941 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca
     6001 aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc
     6061 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa
     6121 cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct
     6181 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattaattc tgtggaatgt
     6241 gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     6301 gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag
     6361 tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat
     6421 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt
     6481 tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt agtgaggagg
     6541 cttttttgga ggcctaggct tttgcaaaaa gctcccggga gcttgtatat ccattttcgg
     6601 atctgatcaa gagacaggat gaggatcgtt tcgc
//
