LOCUS       sequence-487456-        6605 bp    DNA     circular SYN 30-SEP-2025
DEFINITION  Expresses EPB41L4A-AS1 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6605)
  AUTHORS   Monziani A,  Unfried JP, Cvetanovic T, Ulitsky I
  TITLE     EPB41L4A-AS1 long noncoding RNA acts in both cis- and trans-acting 
            transcriptional regulation and controls nucleolar biology
  JOURNAL   bioRxiv 2025.04.08.647803
REFERENCE   2  (bases 1 to 6605)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Sep 30, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "bioRxiv 
            2025.04.08.647803"
COMMENT     Sequence Label: pcDNA3.1(+) EPB41L4A-AS1
FEATURES             Location/Qualifiers
     source          1..6605
                     /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(2196..2213)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2202..2426
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2472..2900
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2559..2578)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2769..2790
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2909..2929)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2914..3243
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3094..3229
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3156..3175
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3310..4104
                     /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(3364..3383)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3974..3993
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4278..4399
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4315..4334)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4369..4388
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4448..4464)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4448..4464)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4461..4483)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4472..4488
                     /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(4496..4502,4503..4520,4521..4526))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4541..4562
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4679..4696)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4850..5438)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4930..4949)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5609..6400,6401..6469))
                     /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     6232..6251
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6470..6574)
                     /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 aagcttggta ccgagctcgg atccactagt ccagtgtggt ggaattctgt
      961 cgggtcccaa aatacgtggt ctaaagtcct ttgggctcct gccgctgcca gcactgatca
     1021 tatgctcttg accctggtcg ggtgggggga tgtctggata cctcctctgt ggaaagaact
     1081 ggccacacat ccgcagaggt gactcgggct gagaacccag gctcctcctc cctttggtga
     1141 cacgcccctc ggtccctcac tggcacttct ccctccggcc acacggcggc gtctcgccat
     1201 agcgcagcgg ccgatggtac agcccgctcc cccctcgcgc tctcggacag tgggtccttc
     1261 cacttgtaga aaagcattgt gggacggaag cctgtccttt cttccttttg gtgcgagctt
     1321 gctgtggttt ttgctctggg tcctctggga tggcgcctgg ctgtggccgc gtggtctctc
     1381 acgcaggggc gccgggcggg ggaacgcggc caccctgagt ctggtgagtc gactgcggcg
     1441 gcctgtgtcc gaagtgtccg gggccgtgaa caagggcagc ggcctggcct caggcctgcg
     1501 ttcccacgtt tggaaacggg gagcttcgtc gatttgtgtt tacatcatcg actatgccag
     1561 ggagttctcc agataagcct ggttttattt tcgtcagtga aaaggcctta ccgtataact
     1621 gactttatgc ttgccctgcc cccgtataaa ataacttaaa agcagcgtgc ctggttacag
     1681 ctgtttccac gtgcggtgct cgtcgggagt gatcacctac cctacaggtg gaagatggat
     1741 gcctgaagtg tagactgctg ctagctgaat accatctggg agcataaagg tgacctgaag
     1801 gatgtccttg gtgaggattt tgaaaatttg atcttcacaa gagttgcctg gatcatttga
     1861 aatttctggg agtctgagga gtactgacat aattacctgc tggagtctgt aaatacacat
     1921 ttaagacagt gaggatgtga ataaatatat taatgcactt tggcatttgt gttttaagtg
     1981 attaactgcc agaaacagct atttctaaaa agttataagg gaggaggggt tcttttttga
     2041 tcagtattca ctgctgtcac cataattaat agccttaaaa tagcttgtgt ttggcccaaa
     2101 gagaaatgta ctttcttcca gtgactcaaa aatcgtggct agaactagac tgcggccgct
     2161 cgagtctaga gggcccgttt aaacccgctg atcagcctcg actgtgcctt ctagttgcca
     2221 gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac
     2281 tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat
     2341 tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca
     2401 tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctctag
     2461 ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     2521 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
     2581 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
     2641 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
     2701 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     2761 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
     2821 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
     2881 acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc
     2941 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg
     3001 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     3061 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
     3121 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
     3181 tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
     3241 aaaaagctcc cgggagcttg tatatccatt ttcggatctg atcaagagac aggatgagga
     3301 tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag
     3361 aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc
     3421 cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg
     3481 aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc
     3541 gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg
     3601 ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct
     3661 gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg
     3721 aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat
     3781 ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc
     3841 atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg
     3901 gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc
     3961 tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct
     4021 gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat
     4081 cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga
     4141 cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct
     4201 tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg
     4261 agttcttcgc ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     4321 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     4381 aactcatcaa tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt
     4441 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
     4501 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
     4561 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     4621 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     4681 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     4741 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     4801 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     4861 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     4921 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     4981 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     5041 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     5101 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     5161 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     5221 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     5281 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     5341 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     5401 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     5461 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     5521 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     5581 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     5641 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     5701 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     5761 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     5821 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     5881 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     5941 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     6001 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     6061 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     6121 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     6181 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     6241 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     6301 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     6361 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     6421 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     6481 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     6541 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     6601 acgtc
//
