LOCUS       sequence-178523-        6585 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mammalian expression of PAFAH1B1 R22 mutant. The protein is 
            truncated and non functional (negative control)..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6585)
  AUTHORS   Josipovic I, Fork C, Preussner J, Prior KK, Iloska D, Vasconez AE, 
            Labocha S, Angioni C, Thomas D, Ferreiros N, Looso M, Pullamsetti 
            SS, Geisslinger G, Steinhilber D, Brandes RP, Leisegang MS
  TITLE     PAFAH1B1 and the lncRNA NONHSAT073641 maintain an angiogenic 
            phenotype in human endothelial cells.
  JOURNAL   Acta Physiol (Oxf). 2016 Apr 28. doi: 10.1111/apha.12700.
  PUBMED    27124368
REFERENCE   2  (bases 1 to 6585)
  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: "Acta 
            Physiol (Oxf). 2016 Apr 28. doi: 10.1111/apha.12700."
COMMENT     Sequence Label: pcDNA3.1+-PAFAH1B1_R22-
FEATURES             Location/Qualifiers
     source          1..6585
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2179..2196)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2185..2409
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2455..2883
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2542..2561)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2752..2773
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2892..2912)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2897..3226
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3077..3212
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3139..3158
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3293..4087
                     /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(3347..3366)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3957..3976
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4261..4382
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4298..4317)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4352..4371
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4431..4447)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4431..4447)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4444..4466)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4455..4471
                     /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(4479..4485,4486..4503,4504..4509))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4524..4545
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4662..4679)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4833..5418)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4913..4932)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5589..6380,6381..6449))
                     /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     6212..6231
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6450..6554)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaactt aagctagctt atggtgctgt cccagagaca atgagatgaa ctaaatcgag
      961 ctatagcaga ttatcttcgt tcaaatggct atgaagaggc atattcagtt tttaaaaagg
     1021 aagctgaatt agatgtgaat gaagaattag ataaaaagta tgctggtctt ttggaaaaaa
     1081 aatggacatc tgttattaga ttacaaaaga aggttatgga attagaatca aagctaaatg
     1141 aagcaaaaga agaatttacg tcaggtggac ctcttggtca gaaacgagac ccaaaagaat
     1201 ggattccccg tccgccagaa aaatatgcat tgagtggtca caggagtcca gtcactcgag
     1261 tcattttcca tcctgtgttc agtgttatgg tctctgcttc agaggatgct acaattaagg
     1321 tgtgggatta tgagactgga gattttgaac gaactcttaa aggacataca gactctgtac
     1381 aggacatttc attcgaccac agcggcaagc ttctggcttc ctgttctgca gatatgacca
     1441 ttaaactatg ggattttcag ggctttgaat gcatcagaac catgcacggc catgaccaca
     1501 atgtttcttc agtagccatc atgcccaatg gagatcatat agtgtctgcc tcaagggata
     1561 aaactataaa aatgtgggaa gtgcaaactg gctactgtgt gaagacattc acaggacaca
     1621 gagaatgggt acgtatggta cggccaaatc aagatggcac tctgatagcc agctgttcca
     1681 atgaccagac tgtgcgtgta tgggtcgtag caacaaagga atgcaaggct gagctccgag
     1741 agcatgagca tgtggtagaa tgcatttcct gggctccaga aagctcatat tcctccatct
     1801 ctgaagcaac aggatctgag actaaaaaaa gtggtaaacc tgggccattc ttgctgtctg
     1861 gatccagaga caagactatt aagatgtggg atgtcagtac tggcatgtgc cttatgaccc
     1921 tcgtgggtca tgataactgg gtacgtggag ttctgttcca ttctgggggg aagtttattt
     1981 tgagttgtgc tgatgacaag accctacgcg tatgggatta caagaacaag cgatgcatga
     2041 agaccctcaa tgcgcatgaa cactttgtta cctccttgga tttccacaag acggcaccct
     2101 atgtcgtcac tggcagcgta gatcaaacag taaaagtgtg ggagtgccgt tgagggcccg
     2161 tttaaacccg ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc
     2221 cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa
     2281 atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg
     2341 ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg
     2401 gctctatggc ttctgaggcg gaaagaacca gctggggctc tagggggtat ccccacgcgc
     2461 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     2521 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     2581 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
     2641 tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc
     2701 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
     2761 tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga
     2821 ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
     2881 attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg
     2941 cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg
     3001 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc
     3061 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
     3121 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt
     3181 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc
     3241 ttgtatatcc attttcggat ctgatcaaga gacaggatga ggatcgtttc gcatgattga
     3301 acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga
     3361 ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg
     3421 gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga
     3481 ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt
     3541 tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct
     3601 gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct
     3661 gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg
     3721 agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca
     3781 ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga
     3841 tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt
     3901 ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt
     3961 ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct
     4021 ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt
     4081 cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca
     4141 cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg
     4201 gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc
     4261 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
     4321 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
     4381 tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg
     4441 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
     4501 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
     4561 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     4621 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     4681 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     4741 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     4801 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     4861 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     4921 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     4981 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     5041 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     5101 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     5161 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     5221 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     5281 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     5341 tccggcaaac aaaccaccgc tggtagcggt ttttttgttt gcaagcagca gattacgcgc
     5401 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     5461 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     5521 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     5581 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     5641 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     5701 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     5761 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     5821 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     5881 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     5941 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     6001 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     6061 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     6121 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     6181 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     6241 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     6301 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     6361 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     6421 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     6481 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     6541 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtc
//
