LOCUS       Exported                5622 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pEF1a-IRES-Neo
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5622)
  AUTHORS   Dejosez M, Levine SS, Frampton GM, Whyte WA, Stratton SA, Barton MC,
            Gunaratne PH, Young RA, Zwaka TP
  TITLE     Ronin/Hcf-1 binds to a hyperconserved enhancer element and regulates
            genes involved in the growth of embryonic stem cells.
  JOURNAL   Genes Dev. 2010 Jul 15. 24(14):1479-84.
  PUBMED    20581084
REFERENCE   2  (bases 1 to 5622)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5622
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        19..1181
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          250..1172
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1129..1149
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     intron          1220..1352
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1335..1354
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        1397..1415
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    1481..2033
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1625..1642)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1852..1871
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             2033..2830
                     /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="MGIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGR
                     PVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLL
                     SSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEH
                     QGLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDI
                     ALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2090..2109)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2700..2719
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    2894..2942
                     /note="synthetic polyadenylation signal"
     rep_origin      3037..3492
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3124..3143)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3334..3355
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3559..3578)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3678..3700
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3738..3756)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3824..3928
                     /gene="bla"
                     /label=AmpR promoter
     CDS             3929..4789
                     /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(4147..4166)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4960..5548
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5449..5468
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ccaattcaag cttcgtgagg ctccggtgcc cgtcagtggg cagagcgcac atcgcccaca
       61 gtccccgaga agttgggggg aggggtcggc aattgaaccg gtgcctagag aaggtggcgc
      121 ggggtaaact gggaaagtga tgtcgtgtac tggctccgcc tttttcccga gggtggggga
      181 gaaccgtata taagtgcagt agtcgccgtg aacgttcttt ttcgcaacgg gtttgccgcc
      241 agaacacagg taagtgccgt gtgtggttcc cgcgggcctg gcctctttac gggttatggc
      301 ccttgcgtgc cttgaattac ttccacctgg ctccagtacg tgattcttga tcccgagctg
      361 gagccagggg cgggccttgc gctttaggag ccccttcgcc tcgtgcttga gttgaggcct
      421 ggcctgggcg ctggggccgc cgcgtgcgaa tctggtggca ccttcgcgcc tgtctcgctg
      481 ctttcgataa gtctctagcc atttaaaatt tttgatgacc tgctgcgacg ctttttttct
      541 ggcaagatag tcttgtaaat gcgggccagg atctgcacac tggtatttcg gtttttgggg
      601 ccgcgggcgg cgacggggcc cgtgcgtccc agcgcacatg ttcggcgagg cggggcctgc
      661 gagcgcggcc accgagaatc ggacgggggt agtctcaagc tggccggcct gctctggtgc
      721 ctggcctcgc gccgccgtgt atcgccccgc cctgggcggc aaggctggcc cggtcggcac
      781 cagttgcgtg agcggaaaga tggccgcttc ccggccctgc tccagggggc tcaaaatgga
      841 ggacgcggcg ctcgggagag cgggcgggtg agtcacccac acaaaggaaa ggggcctttc
      901 cgtcctcagc cgtcgcttca tgtgactcca cggagtaccg ggcgccgtcc aggcacctcg
      961 attagttctg gagcttttgg agtacgtcgt ctttaggttg gggggagggg ttttatgcga
     1021 tggagtttcc ccacactgag tgggtggaga ctgaagttag gccagcttgg cacttgatgt
     1081 aattctcctt ggaatttgcc ctttttgagt ttggatcttg gttcattctc aagcctcaga
     1141 cagtggttca aagttttttt cttccatttc aggtgtcgtg aacacgggct gcagaagttg
     1201 gtcgtgaggc actgggcagg taagtatcaa ggttacaaga caggtttaag gagaccaata
     1261 gaaactgggc ttgtcgagac agagaagact cttgcgtttc tgataggcac ctattggtct
     1321 tactgacatc cactttgcct ttctctccac aggtgtccac tcccagttca attacagctc
     1381 ttaaggctag agtacttaat acgactcact ataggctagc ctcgagatcc tagagtcgac
     1441 ccgggcggcc gctctagccc aattccgccc ccccccccta acgttactgg ccgaagccgc
     1501 ttggaataag gccggtgtgc gtttgtctat atgttatttt ccaccatatt gccgtctttt
     1561 ggcaatgtga gggcccggaa acctggccct gtcttcttga cgagcattcc taggggtctt
     1621 tcccctctcg ccaaaggaat gcaaggtctg ttgaatgtcg tgaaggaagc agttcctctg
     1681 gaagcttctt gaagacaaac aacgtctgta gcgacccttt gcaggcagcg gaacccccca
     1741 cctggcgaca ggtgcctctg cggccaaaag ccacgtgtat aagatacacc tgcaaaggcg
     1801 gcacaacccc agtgccacgt tgtgagttgg atagttgtgg aaagagtcaa atggctctcc
     1861 tcaagcgtat tcaacaaggg gctgaaggat gcccagaagg taccccattg tatgggatct
     1921 gatctggggc ctcggtgcac atgctttaca tgtgtttagt cgaggttaaa aaaacgtcta
     1981 ggccccccga accacgggga cgtggttttc ctttgaaaaa cacgatgata atatgggcat
     2041 tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta
     2101 tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca
     2161 ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga
     2221 cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga
     2281 cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct
     2341 cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg
     2401 gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga
     2461 gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca
     2521 tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga
     2581 ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg
     2641 cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc
     2701 gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt
     2761 gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga
     2821 gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca
     2881 tcacgatggc cgcaataaaa tatctttatt ttcattacat ctgtgtgttg gttttttgtg
     2941 tgaatcgata gcgataagga tccgggctgg cgtaatagcg aagaggcccg caccgatcgc
     3001 ccttcccaac agttgcgcag cctgaatggc gaatggacgc gccctgtagc ggcgcattaa
     3061 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc
     3121 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag
     3181 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca
     3241 aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc
     3301 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa
     3361 cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct
     3421 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa
     3481 cgcttacaat ttcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg
     3541 catatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc agccccgaca
     3601 cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag
     3661 acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa
     3721 acgcgcgaga cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat
     3781 aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     3841 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     3901 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
     3961 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
     4021 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
     4081 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
     4141 agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg
     4201 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
     4261 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
     4321 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
     4381 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
     4441 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact
     4501 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
     4561 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
     4621 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
     4681 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     4741 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
     4801 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     4861 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     4921 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     4981 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     5041 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     5101 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     5161 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     5221 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     5281 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     5341 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     5401 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     5461 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     5521 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     5581 ggccttttgc tggccttttg ctcacatggc tcgacagatc cc
//
