LOCUS       sequence-289645-        6470 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian Expression of HsNanos1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6470)
  AUTHORS   Bhandari D, Raisch T, Weichenrieder O, Jonas S, Izaurralde E
  TITLE     Structural basis for the Nanos-mediated recruitment of the CCR4-NOT 
            complex and translational repression.
  JOURNAL   Genes Dev. 2014 Apr 15;28(8):888-901. doi: 10.1101/gad.237289.113.
  PUBMED    24736845
REFERENCE   2  (bases 1 to 6470)
  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"; doi: 
            "10.1101/gad.237289"; journalName: "Genes Dev"; date: "2014-04-15- 
            15"; volume: "28"; issue: "8"; pages: "888-901"
COMMENT     Sequence Label: pCIneo-lambdaN-HA-HsNanos1_P
FEATURES             Location/Qualifiers
     source          1..6470
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        138..517
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        518..729
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     680..700
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     726..750
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          890..1022
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1005..1024
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        1067..1085
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      1098..1107
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1104..1169
                     /codon_start=1
                     /product="N-terminal RNA-binding domain from the lambda 
                     bacteriophage antiterminator protein N (Baron-Benhamou et 
                     al., 2004)"
                     /label=lambda N peptide
                     /translation="MDAQTRRRERRAEKQAQWKAAN"
     CDS             1194..1220
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    2174..2295
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2185..2204)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2239..2258
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      2481..2936
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2568..2587)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2778..2799
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3053..3410
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3053..3244
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3076..3096)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3261..3396
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3461..4255
                     /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(3515..3534)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4125..4144
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4319..4367
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(4408..4427)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4527..4549
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4587..4605)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4673..4777
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4778..4846,4847..5638)
                     /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(4996..5015)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5809..6397
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6298..6317
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tcaatattgg ccattagcca tattattcat tggttatata gcataaatca atattggcta
       61 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
      121 aatatgaccg ccatgttggc attgattatt gactagttat taatagtaat caattacggg
      181 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      241 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      301 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      361 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
      421 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
      481 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
      541 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      601 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactg
      661 cgatcgcccg ccccgttgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata
      721 agcagagctc gtttagtgaa ccgtcagatc actagaagct ttattgcggt agtttatcac
      781 agttaaattg ctaacgcagt cagtgcttct gacacaacag tctcgaactt aagctgcagt
      841 gactctctta aggtagcctt gcagaagttg gtcgtgaggc actgggcagg taagtatcaa
      901 ggttacaaga caggtttaag gagaccaata gaaactgggc ttgtcgagac agagaagact
      961 cttgcgtttc tgataggcac ctattggtct tactgacatc cactttgcct ttctctccac
     1021 aggtgtccac tcccagttca attacagctc ttaaggctag agtacttaat acgactcact
     1081 ataggctagc aattgcggcc accatggacg cacaaacacg acgacgtgag cgtcgcgctg
     1141 agaaacaagc tcaatggaaa gctgcaaacg gtggatctcc tccacatatg gcttacccat
     1201 acgatgttcc agattacgct cctccactcg agatggaagc cttcccatgg gcccccagat
     1261 cccccagaag aggcagagca cctcctccca tggccctggt gccaagcgcc agatatgtgt
     1321 ctgcccctgg acctgcccac ccccagcctt tcagcagctg gaacgattac ctgggcctgg
     1381 ccaccctgat caccaaggcc gtggatggcg agcccagatt cggatgtgcc agaggcggaa
     1441 atggcggcgg aggcagccct cctagtagca gcagcagctc ctgttgcagc cctcacacag
     1501 gcgctggacc tggcgctctg ggacctgcac tgggcccacc tgactacgac gaggacgacg
     1561 acgatgacag cgacgagcct ggcagcagag gcagatacct gggctctgcc ctggaactgc
     1621 gggctctgga actgtgtgcc ggacctgctg aagccggcct gctggaagag agattcgccg
     1681 agctgagccc cttcgccggc agagctgcag cagtgctgct gggatgtgct cctgctgccg
     1741 cagccgccgc tacaacaaca tctgaggcca cccctcggga agagagggct cctgcttggg
     1801 ccgctgaacc tagactgcat gctgcctctg gcgccgctgc tgcaagactg ctgaaacccg
     1861 aactgcaagt gtgcgtgttc tgccggaaca acaaagaagc tatggccctg tacaccaccc
     1921 acatcctgaa gggccccgac ggcagagtgc tgtgtcccgt gctgcggaga tacacatgcc
     1981 ctctgtgtgg cgccagcggc gacaatgccc acaccatcaa gtactgcccc ctgagcaagg
     2041 tgccccctcc accagctaga ccccctccaa gatctgccag agatggcccc cctggcaaga
     2101 agctgagata atctagagtc gacccgggcg gccgcttccc tttagtgagg gttaatgctt
     2161 cgagcagaca tgataagata cattgatgag tttggacaaa ccacaactag aatgcagtga
     2221 aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac cattataagc
     2281 tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt tcagggggag
     2341 atgtgggagg ttttttaaag caagtaaaac ctctacaaat gtggtaaaat ccgataagga
     2401 tcgatccggg ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc
     2461 gcagcctgaa tggcgaatgg acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     2521 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     2581 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     2641 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
     2701 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     2761 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
     2821 ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga
     2881 gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgctta caatttcctg
     2941 atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatacg cggatctgcg
     3001 cagcaccatg gcctgaaata acctctgaaa gaggaacttg gttaggtacc ttctgaggcg
     3061 gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag
     3121 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc
     3181 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
     3241 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     3301 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc
     3361 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagcttgatt
     3421 cttctgacac aacagtctcg aacttaaggc tagagccacc atgattgaac aagatggatt
     3481 gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca
     3541 gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct
     3601 ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct
     3661 atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc
     3721 gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct
     3781 tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga
     3841 tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg
     3901 gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc
     3961 agccgaactg ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac
     4021 ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat
     4081 cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga
     4141 tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc
     4201 cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg
     4261 actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg atggccgcaa
     4321 taaaatatct ttattttcat tacatctgtg tgttggtttt ttgtgtgaat cgatagcgat
     4381 aaggatccgc gtatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca
     4441 gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc
     4501 cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc
     4561 atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt
     4621 catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac
     4681 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     4741 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
     4801 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
     4861 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
     4921 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
     4981 cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca
     5041 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
     5101 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
     5161 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
     5221 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     5281 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt
     5341 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
     5401 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
     5461 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg
     5521 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
     5581 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
     5641 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     5701 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     5761 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     5821 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     5881 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     5941 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     6001 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     6061 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     6121 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     6181 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     6241 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     6301 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     6361 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     6421 acggttcctg gccttttgct ggccttttgc tcacatggct cgacagatct
//
