LOCUS       sequence-217932-        6715 bp    DNA     circular SYN 15-APR-2025
DEFINITION  transient/stable overexpression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6715)
  AUTHORS   Niewiadomski P, Proszynski TJ
  TITLE     LL5-beta interacts with uncharacterised protein 2410002F23Rik 
            (LL5BIP)
  JOURNAL   Matters v. 2, n. 11  (Nov 17, 2016)
REFERENCE   2  (bases 1 to 6715)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Matters v. 
            2, n. 11  (Nov 17, 2016)"
COMMENT     Sequence Label: pcDNA3.1(+)/Puro/FA/LL5BIP-GFP
FEATURES             Location/Qualifiers
     source          1..6715
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        8..387
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        388..591
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     542..562
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     636..655
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        636..654
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             join(1583..1585,1586..1588,1589..2302)
                     /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             join(1583..1585,1586..1588,1589..2299)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1628..1649)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1889..1908)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2236..2257
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2334..2351)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2340..2564
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2610..3038
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2697..2716)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2907..2928
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3047..3067)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3052..3381
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3232..3367
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3294..3313
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3402..4001
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(3402..3421)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3898..3918
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    4161..4282
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4198..4217)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4252..4271
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4331..4347)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4331..4347)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4344..4366)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4355..4371
                     /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(4379..4385,4386..4403,4404..4409))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4424..4445
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4562..4579)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4733..5321)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4813..4832)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5492..6283,6284..6352))
                     /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     6115..6134
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6353..6457)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6532..6551)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 acgcgttgac attgattatt gactagttat taatagtaat caattacggg gtcattagtt
       61 catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga
      121 ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca
      181 atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca
      241 gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg
      301 cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc
      361 tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt
      421 ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt
      481 ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg
      541 acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc tctctggcta
      601 actagagaac ccactgctta ctggcttatc gaaattaata cgactcacta tagggagacc
      661 caagctggct agcgtttaaa cttaagcttg gtaccgagct cggatccacg gccggccaac
      721 catggctgag agagtgctgg tgcccaccca gataggccgg ggggaccgct actacacata
      781 cacggagttg ttggctatct cacggcgttt caagcagaac cccaatgagc tcatggtcac
      841 ctggatcctg cgggtgtatg accagggagg ccctgccctg tccctgaatt ctggggagct
      901 ggggctgctg ggtgacctca cccatgatgc catctttaac taccgctgca aggccctgcg
      961 gggggctggc tgccagactc tcctgagctg gctgctgcag gcctggcgtc agcgttggga
     1021 atcctccctg cattttgagg ccaccgagtt acccttcagg ccctggacca ccatggagga
     1081 aggcatccag ttggtgcgtg agctgggcat gattgagtgg atctaccttg acccagaagg
     1141 gcccgtggac ctggccccag aggatgtggc cttcactcaa ggcctgcagc ggcgcctgct
     1201 cacagcagcc ccctctgagc tgcggctttc actggtcagc ctgctggtac gtggcatgac
     1261 agtgttggag gctgtgatgg agatccagac tattgctgac gtgggtctgc tctggcgcca
     1321 gagccatcca ggccgcacca agctcatgtt ggggcccaac ccaactcgta aggacctcct
     1381 aggctggctg ctcagccacg gtgtaccccg ggagcaagta gacagacagc ccaccaaggt
     1441 actcctagaa ctctacatca aagaagccaa gcgcagtcgc ggccacccca actatgggct
     1501 gaacgaggag cagcccccac caccccctta ctccgaccag gcctgtgggg aagagcaacc
     1561 ggtgcgtcat gacggcgcgc ctatggtgag caagggcgag gagctgttca ccggggtggt
     1621 gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga
     1681 gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa
     1741 gctgcccgtg ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag
     1801 ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta
     1861 cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt
     1921 gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga
     1981 ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat
     2041 catggccgac aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga
     2101 ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc
     2161 cgtgctgctg cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa
     2221 cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg
     2281 catggacgag ctgtacaagt aatctagagg gcccgtttaa acccgctgat cagcctcgac
     2341 tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct
     2401 ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct
     2461 gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg
     2521 ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag
     2581 aaccagctgg ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc
     2641 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc
     2701 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
     2761 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
     2821 tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
     2881 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
     2941 ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt
     3001 aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag
     3061 ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     3121 aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     3181 agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc
     3241 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
     3301 gcagaggccg aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt
     3361 ggaggcctag gcttttgcaa aaagctcccg gggccgccac catgaccgag tacaagccca
     3421 cggtgcgcct cgccacccgc gacgacgtcc cccgggccgt acgcaccctc gccgccgcgt
     3481 tcgccgacta ccccgccacg cgccacaccg tcgacccgga ccgccacatc gagcgggtca
     3541 ccgagctgca agaactcttc ctcacgcgcg tcgggctcga catcggcaag gtgtgggtcg
     3601 cggacgacgg cgccgcggtg gcggtctgga ccacgccgga gagcgtcgaa gcgggggcgg
     3661 tgttcgccga gatcggcccg cgcatggccg agttgagcgg ttcccggctg gccgcgcagc
     3721 aacagatgga aggcctcctg gcgccgcacc ggcccaagga gcccgcgtgg ttcctggcca
     3781 ccgtcggcgt ctcgcccgac caccagggca agggtctggg cagcgccgtc gtgctccccg
     3841 gagtggaggc ggccgagcgc gccggggtgc ccgccttcct ggagacctcc gcgccccgca
     3901 acctcccctt ctacgagcgg ctcggcttca ccgtcaccgc cgacgtcgag gtgcccgaag
     3961 gaccgcgcac ctggtgcatg acccgcaagc ccggtgcctg attcgaaatg accgaccaag
     4021 cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg
     4081 gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc
     4141 tggagttctt cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca
     4201 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
     4261 ccaaactcat caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg
     4321 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
     4381 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
     4441 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     4501 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     4561 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     4621 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     4681 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     4741 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     4801 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     4861 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     4921 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     4981 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     5041 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     5101 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     5161 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     5221 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     5281 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     5341 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     5401 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     5461 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     5521 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     5581 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     5641 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     5701 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     5761 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     5821 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     5881 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     5941 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     6001 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     6061 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     6121 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     6181 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     6241 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     6301 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     6361 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     6421 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     6481 ctgacgtcga cggatcggga gatctcccga tcccctatgg tgcactctca gtacaatctg
     6541 ctctgatgcc gcatagttaa gccagtatct gctccctgct tgtgtgttgg aggtcgctga
     6601 gtagtgcgcg agcaaaattt aagctacaac aaggcaaggc ttgaccgaca attgcatgaa
     6661 gaatctgctt agggttaggc gttttgcgct gcttcgcgat gtacgggcca gatat
//
