LOCUS       sequence-420602-        7065 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 7065)
  AUTHORS   Itakura E, Kishi C, Inoue K, Mizushima N
  TITLE     Beclin 1 forms two distinct phosphatidylinositol 3-kinase complexes 
            with mammalian Atg14 and UVRAG.
  JOURNAL   Mol Biol Cell. 2008 Dec . 19(12):5360-72.
  PUBMED    18843052
REFERENCE   2  (bases 1 to 7065)
  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: "Mol Biol 
            Cell. 2008 Dec . 19(12):5360-72."
COMMENT     Sequence Label: pCI-neo-HA-hAtg14
FEATURES             Location/Qualifiers
     source          1..7065
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             43..132
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAGYPYDVPDYAGSYPYDVPDYA"
     polyA_signal    1699..1820
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1710..1729)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1764..1783
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      2006..2461
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2093..2112)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2303..2324
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2578..2935
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2578..2769
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2601..2621)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2786..2921
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2986..3780
                     /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(3040..3059)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3650..3669
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3844..3892
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(3933..3952)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4052..4074
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4112..4130)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4198..4302
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4303..4371,4372..5163)
                     /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     complement(4521..4540)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5334..5922
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5823..5842
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        6137..6516
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6517..6728
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6679..6699
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     6725..6749
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          6889..7021
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     7004..7023
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
ORIGIN
        1 taatacgact cactataggc tagcctcgag agaatgatct tttacccata cgatgttcct
       61 gactatgcgg gctatcccta tgacgtcccg gactatgcag gatcctatcc atatgacgtt
      121 ccagattacg ctgctcagtg cactcgagaa ttggggaatt ccatggcgtc tcccagtggg
      181 aagggagccc gggcgctgga ggctcctggc tgcgggcccc ggccgctcgc ccgggacctg
      241 gtggactccg tggacgatgc ggaggggctg tacgtggctg tggagcgctg cccgctgtgc
      301 aacactaccc gccggcggct gacctgcgcc aaatgcgttc agagcggcga tttcgtctac
      361 ttcgacggcc gcgaccggga gaggtttatc gacaagaagg aaaggttaag ccgacttaag
      421 agcaagcaag aagaatttca gaaagaagtg ttaaaagcta tggaaggaaa atggataaca
      481 gatcagttga gatggaaaat aatgtcctgc aagatgagga ttgaacagtt aaaacaaaca
      541 atatgtaaag gaaatgaaga aatggagaaa aattctgaag gccttctcaa aaccaaggaa
      601 aagaatcaga agctttacag tcgagcacaa cggcaccaag agaaaaagga gaagattcag
      661 aggcataatc gcaaacttgg tgacctggta gaaaaaaaga ccattgactt aagaagtcat
      721 tatgagcgtc tggcaaatct tcgacgatcc catatattag agctcacctc tgtcattttt
      781 ccaatcgagg aagtaaagac gggtgtgaga gaccccgcag atgtgtcttc agagagtgac
      841 agtgccatga cctccagcac tgtgagcaag cttgctgaag cccggaggac aacttacctc
      901 tcaggacgat gggtctgtga cgatcacaac ggagacacca gcattagcat tacagggcct
      961 tggattagcc tccctaacaa tggggactac tctgcctact acagctgggt ggaggagaag
     1021 aaaacaaccc aggggcctga catggagcag agtaaccctg cctacaccat cagtgctgcg
     1081 ctgtgctatg caactcagct ggtcaacatt ctgtctcata tacttgatgt aaatcttccc
     1141 aaaaagctct gcaacagtga attttgtggc gaaaatctaa gcaagcagaa atttactcga
     1201 gcagtgaaga aactgaatgc aaatattctt tacctttgtt tttctcagca tgtaaattta
     1261 gatcaattac aaccactgca taccctcagg aatctaatgt acctggtcag tccaagctct
     1321 gaacacctag gcaggtcagg gccctttgaa gtacgagcag accttgagga gtccatggaa
     1381 tttgtggatc ccggagttgc tggagaatca gatgagagcg gagatgagcg cgtcagcgat
     1441 gaagaaaccg acctgggcac agactgggag aacttgccta gtccccggtt ttgtgatatc
     1501 ccttcccagt ctgtggaagt ctcccagagt cagagcaccc aggcgtcccc acccatcgcg
     1561 agcagcagtg caggtgggat gatctcctct gcagcagcct cggtgacctc ctggtttaaa
     1621 gcttacactg gacaccgtta agaattcccc gggcggccgc ttccctttag tgagggttaa
     1681 tgcttcgagc agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc
     1741 agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta
     1801 taagctgcaa taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg
     1861 gggagatgtg ggaggttttt taaagcaagt aaaacctcta caaatgtggt aaaatccgat
     1921 aaggatcgat ccgggctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca
     1981 gttgcgcagc ctgaatggcg aatggacgcg ccctgtagcg gcgcattaag cgcggcgggt
     2041 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
     2101 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
     2161 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
     2221 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     2281 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
     2341 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
     2401 aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac gcttacaatt
     2461 tcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atacgcggat
     2521 ctgcgcagca ccatggcctg aaataacctc tgaaagagga acttggttag gtaccttctg
     2581 aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc
     2641 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa
     2701 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     2761 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
     2821 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc
     2881 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct
     2941 tgattcttct gacacaacag tctcgaactt aaggctagag ccaccatgat tgaacaagat
     3001 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca
     3061 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg
     3121 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg
     3181 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact
     3241 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct
     3301 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg
     3361 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt
     3421 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc
     3481 gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc
     3541 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga
     3601 ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc
     3661 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt
     3721 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga
     3781 gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgatggc
     3841 cgcaataaaa tatctttatt ttcattacat ctgtgtgttg gttttttgtg tgaatcgata
     3901 gcgataagga tccgcgtatg gtgcactctc agtacaatct gctctgatgc cgcatagtta
     3961 agccagcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg
     4021 gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca
     4081 ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt tttataggtt
     4141 aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc
     4201 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     4261 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
     4321 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
     4381 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
     4441 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
     4501 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa
     4561 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
     4621 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
     4681 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
     4741 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
     4801 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca
     4861 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
     4921 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
     4981 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
     5041 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
     5101 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
     5161 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
     5221 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     5281 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     5341 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     5401 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     5461 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
     5521 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     5581 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     5641 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     5701 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     5761 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     5821 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     5881 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     5941 tttttacggt tcctggcctt ttgctggcct tttgctcaca tggctcgaca gatcgatctt
     6001 caatattggc cattagccat attattcatt ggttatatag cataaatcaa tattggctat
     6061 tggccattgc atacgttgta tctatatcat aatatgtaca tttatattgg ctcatgtcca
     6121 atatgaccgc catgttggca ttgattattg actagttatt aatagtaatc aattacgggg
     6181 tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg
     6241 cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata
     6301 gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc
     6361 cacttggcag tacatcaagt gtatcatatg ccaagtccgc cccctattga cgtcaatgac
     6421 ggtaaatggc ccgcctggca ttatgcccag tacatgacct tacgggactt tcctacttgg
     6481 cagtacatct acgtattagt catcgctatt accatggtga tgcggttttg gcagtacacc
     6541 aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc
     6601 aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactgc
     6661 gatcgcccgc cccgttgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa
     6721 gcagagctcg tttagtgaac cgtcagatca ctagaagctt tattgcggta gtttatcaca
     6781 gttaaattgc taacgcagtc agtgcttctg acacaacagt ctcgaactta agctgcagtg
     6841 actctcttaa ggtagccttg cagaagttgg tcgtgaggca ctgggcaggt aagtatcaag
     6901 gttacaagac aggtttaagg agaccaatag aaactgggct tgtcgagaca gagaagactc
     6961 ttgcgtttct gataggcacc tattggtctt actgacatcc actttgcctt tctctccaca
     7021 ggtgtccact cccagttcaa ttacagctct taaggctaga gtact
//
