LOCUS       sequence-197244-        8117 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 8117)
  AUTHORS   Erdmann KS, Mao Y, McCrea HJ, Zoncu R, Lee S, Paradise S, Modregger 
            J, Biemesderfer D, Toomre D, De Camilli P
  TITLE     A role of the Lowe syndrome protein OCRL in early steps of the 
            endocytic pathway.
  JOURNAL   Dev Cell. 2007 Sep . 13(3):377-90.
  PUBMED    17765681
REFERENCE   2  (bases 1 to 8117)
  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: "Dev Cell. 
            2007 Sep . 13(3):377-90."
COMMENT     Sequence Label: pcDNA3-HA-human OCRL
FEATURES             Location/Qualifiers
     source          1..8117
                     /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"
     CDS             673..699
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     promoter        complement(3436..3454)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3437..3454)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3474..3491)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3480..3704
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3750..4178
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3837..3856)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4047..4068
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4187..4207)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4192..4521
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4372..4507
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4434..4453
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4588..5382
                     /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(4642..4661)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5252..5271
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5556..5677
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5593..5612)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5647..5666
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5726..5742)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5726..5742)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5739..5761)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5750..5766
                     /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(5774..5780,5781..5798,5799..5804))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5819..5840
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5957..5974)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6128..6716)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6208..6227)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6887..7678,7679..7747))
                     /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     7510..7529
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7748..7852)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7927..7946)
                     /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 acccaagctt
      661 ggtaccatgg cgtacccata cgacgtccca gactacgcgg gatccgagcc gccgctcccg
      721 gtcggagccc agccgcttgc cactgtcgag ggtatggaga tgaagggtcc tctccgggag
      781 ccctgcgccc tgaccctagc ccagaggaac gggcaatatg agttaataat ccagttgcat
      841 gagaaggaac agcatgttca agatatcatt cctataaata gccacttcag atgtgttcaa
      901 gaagcagaag aaactctttt gattgacata gcttctaaca gtggctgcaa aattcgggtt
      961 cagggggact ggatcagaga gcgccgcttt gaaatccctg atgaggaaca ctgtttgaag
     1021 ttcctctcag ctgtccttgc tgctcagaaa gctcagtcac agcttcttgt tccagagcaa
     1081 aaggactcat ctagctggta ccagaaatta gacactaagg acaaaccttc tgttttttca
     1141 gggcttcttg gatttgaaga caatttttct tctatgaatt tggacaagaa aataaattca
     1201 caaaatcagc ctactgggat tcatcgggaa cccccacctc cacccttttc agtgaataaa
     1261 atgcttccac gtgaaaaaga agcttctaac aaggagcagc ccaaagtgac caacaccatg
     1321 cggaagctct ttgtaccaaa tacccaatct gggcagcggg agggtctcat caaacatatc
     1381 ctggcaaagc gagagaaaga atatgtcaac attcagactt tcagattttt tgttggaact
     1441 tggaatgtga atggccagtc tccagatagc gggttagaac cttggctgaa ctgtgatccc
     1501 aatcctcctg atatctactg cattggattc caagaactgg acttgagcac agaagccttc
     1561 ttctactttg aatctgtgaa ggaacaagaa tggtccatgg ctgtagagag aggtttgcat
     1621 tccaaagcca agtataagaa agttcaactg gtgcgccttg ttgggatgat gcttcttata
     1681 tttgccagaa aggatcagtg tcgatacatt cgtgatattg ctacagaaac agttggaact
     1741 ggaatcatgg ggaaaatggg aaacaaaggt ggggtagctg tgagatttgt atttcacaac
     1801 accacctttt gcattgtcaa ttcccatctg gctgcacacg tggaggactt tgagagaagg
     1861 aatcaagatt ataaggacat ttgtgcgaga atgagttttg tggtcccaaa tcagaccctc
     1921 ccgcagttga acatcatgaa acatgaggtt gtcatttggt tgggagattt gaattataga
     1981 ctttgcatgc ctgatgccaa tgaggtgaaa agtcttatta ataagaaaga ccttcagaga
     2041 ctcttgaaat tcgaccagct aaatattcag cgcacacaga aaaaagcttt tgttgacttc
     2101 aatgaagggg aaatcaagtt catccccact tataagtatg actctaaaac agaccggtgg
     2161 gattccagtg ggaaatgccg ggttccagcc tggtgtgacc gaattctttg gagaggaaca
     2221 aatgttaatc agcttaatta tcggagtcac atggaactga aaaccagcga ccacaagcct
     2281 gttagcgccc tcttccatat tggggtgaag gttgtggatg aacgaaggta ccggaaagtc
     2341 tttgaagata gtgtacgcat catggacaga atggaaaatg acttccttcc ttccttagaa
     2401 ctcagcagga gggagtttgt gtttgaaaat gtgaagtttc ggcaactaca aaaggagaag
     2461 ttccagatca gcaacaatgg acaggttccc tgccattttt ctttcatccc taaacttaat
     2521 gacagccagt actgcaagcc atggcttcgg gctgaacctt ttgagggcta cttggagcca
     2581 aatgagacag tggacatttc tcttgatgtg tatgtcagca aagactctgt aaccatcctg
     2641 aactcgggag aagataagat tgaagatatt ctcgtccttc acctggatcg aggcaaagat
     2701 tacttcttga ctatcagtgg aaattacctc ccaagttgtt ttggcacatc cttagaggct
     2761 ctgtgccgta tgaaaagacc aatccgagaa gttcctgtta ccaaactcat agacttggaa
     2821 gaagacagct tcctagaaaa ggagaaatcc cttctgcaaa tggttccttt ggatgaaggt
     2881 gccagtgaga gaccccttca ggttcccaag gagatctggc ttctagtaga tcacctattc
     2941 aaatacgcct gtcaccagga ggacctgttc cagacccctg gaatgcagga agagctccag
     3001 cagatcattg attgtctgga taccagcatt cctgagacaa tccctggcag caaccactct
     3061 gtggctgaag cactgctcat tttcttggaa gccctgccag agccagtcat ctgttacgag
     3121 ctgtatcagc gatgtcttga ctctgcttat gatccccgga tctgccgaca ggtgatctcc
     3181 cagcttccga gatgccatag aaatgttttc cgttacttga tggcattcct tcgagaactc
     3241 ttaaaattct ctgaatacaa tagcgtcaat gccaacatga tcgctactct cttcactagt
     3301 cttctcctga ggcctccacc caaccttatg gcaagacaga ctccaagtga ccgccagcgt
     3361 gctattcagt tccttctggg ctttctgctt gggagcgaag aagactagct cgagcatgca
     3421 tctagagggc cctattctat agtgtcacct aaatgctaga gctcgctgat cagcctcgac
     3481 tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct
     3541 ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct
     3601 gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg
     3661 ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag
     3721 aaccagctgg ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc
     3781 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc
     3841 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
     3901 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
     3961 tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
     4021 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
     4081 ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt
     4141 aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag
     4201 ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     4261 aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     4321 agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc
     4381 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
     4441 gcagaggccg aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt
     4501 ggaggcctag gcttttgcaa aaagctcccg ggagcttgta tatccatttt cggatctgat
     4561 caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct
     4621 ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc
     4681 tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc
     4741 gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc gtggctggcc
     4801 acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg
     4861 ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag
     4921 aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc
     4981 ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt
     5041 cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc
     5101 gccaggctca aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc
     5161 tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg
     5221 ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag
     5281 cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg
     5341 cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg
     5401 aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct
     5461 tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc
     5521 gcggggatct catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg
     5581 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     5641 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct
     5701 ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
     5761 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
     5821 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     5881 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     5941 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     6001 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     6061 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     6121 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     6181 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     6241 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     6301 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     6361 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     6421 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     6481 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     6541 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     6601 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     6661 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     6721 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     6781 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     6841 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     6901 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     6961 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     7021 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     7081 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     7141 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     7201 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     7261 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     7321 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     7381 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     7441 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     7501 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     7561 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     7621 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     7681 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     7741 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     7801 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     7861 cccgaaaagt gccacctgac gtcgacggat cgggagatct cccgatcccc tatggtgcac
     7921 tctcagtaca atctgctctg atgccgcata gttaagccag tatctgctcc ctgcttgtgt
     7981 gttggaggtc gctgagtagt gcgcgagcaa aatttaagct acaacaaggc aaggcttgac
     8041 cgacaattgc atgaagaatc tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg
     8101 ggccagatat acgcgtt
//
