LOCUS       sequence-182377-        6584 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 6584)
  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 6584)
  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..6584
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     629..648
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        629..647
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1941..1958)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1947..2171
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2217..2645
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2304..2323)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2514..2535
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2654..2674)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2659..2988
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2839..2974
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2901..2920
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3055..3849
                     /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(3109..3128)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3719..3738
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4023..4144
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4060..4079)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4114..4133
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4193..4209)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4193..4209)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4206..4228)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4217..4233
                     /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(4241..4247,4248..4265,4266..4271))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4286..4307
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4424..4441)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4595..5183)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4675..4694)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5354..6145,6146..6214))
                     /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     5977..5996
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6215..6319)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6394..6413)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag
      601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctg
      661 gctagcgttt aaacttaagc ttatggtgct gtcccagaga caatgagatg aactaaatcg
      721 agctatagca gattatcttc gttcaaatgg ctatgaagag gcatattcag tttttaaaaa
      781 ggaagctgaa ttagatgtga atgaagaatt agataaaaag tatgctggtc ttttggaaaa
      841 aaaatggaca tctgttatta gattacaaaa gaaggttatg gaattagaat caaagctaaa
      901 tgaagcaaaa gaagaattta cgtcaggtgg acctcttggt cagaaacgag acccaaaaga
      961 atggattccc cgtccgccag aaaaatatgc attgagtggt cacaggagtc cagtcactcg
     1021 agtcattttc catcctgtgt tcagtgttat ggtctctgct tcagaggatg ctacaattaa
     1081 ggtgtgggat tatgagactg gagattttga acgaactctt aaaggacata cagactctgt
     1141 acaggacatt tcattcgacc acagcggcaa gcttctggct tcctgttctg cagatatgac
     1201 cattaaacta tgggattttc agggctttga atgcatcaga accatgcacg gccatgacca
     1261 caatgtttct tcagtagcca tcatgcccaa tggagatcat atagtgtctg cctcaaggga
     1321 taaaactata aaaatgtggg aagtgcaaac tggctactgt gtgaagacat tcacaggaca
     1381 cagagaatgg gtacgtatgg tacggccaaa tcaagatggc actctgatag ccagctgttc
     1441 caatgaccag actgtgcgtg tatgggtcgt agcaacaaag gaatgcaagg ctgagctccg
     1501 agagcatgag catgtggtag aatgcatttc ctgggctcca gaaagctcat attcctccat
     1561 ctctgaagca acaggatctg agactaaaaa aagtggtaaa cctgggccat tcttgctgtc
     1621 tggatccaga gacaagacta ttaagatgtg ggatgtcagt actggcatgt gccttatgac
     1681 cctcgtgggt catgataact gggtacgtgg agttctgttc cattctgggg ggaagtttat
     1741 tttgagttgt gctgatgaca agaccctacg cgtatgggat tacaagaaca agcgatgcat
     1801 gaagaccctc aatgcgcatg aacactttgt tacctccttg gatttccaca agacggcacc
     1861 ctatgtcgtc actggcagcg tagatcaaac agtaaaagtg tgggagtgcc gttgagggcc
     1921 cgtttaaacc cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg
     1981 cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata
     2041 aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt
     2101 ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt
     2161 gggctctatg gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc
     2221 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     2281 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     2341 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
     2401 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc
     2461 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     2521 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg
     2581 gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     2641 gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
     2701 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
     2761 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
     2821 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     2881 catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta
     2941 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga
     3001 gcttgtatat ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt
     3061 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat
     3121 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag
     3181 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac
     3241 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac
     3301 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc
     3361 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg
     3421 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag
     3481 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat
     3541 caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag
     3601 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc
     3661 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg
     3721 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg
     3781 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag
     3841 ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat
     3901 cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc
     3961 gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc
     4021 ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     4081 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     4141 cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc
     4201 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
     4261 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
     4321 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     4381 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     4441 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     4501 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     4561 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     4621 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     4681 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     4741 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     4801 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     4861 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     4921 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     4981 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     5041 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     5101 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     5161 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     5221 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     5281 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     5341 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     5401 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     5461 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     5521 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     5581 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     5641 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     5701 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     5761 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     5821 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     5881 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     5941 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     6001 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     6061 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     6121 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     6181 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     6241 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     6301 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc gacggatcgg
     6361 gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt
     6421 aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg cgagcaaaat
     6481 ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc ttagggttag
     6541 gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgtt
//
