LOCUS       Exported                6519 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    Flag-hPIASx beta
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6519)
  AUTHORS   Arora T, Liu B, He H, Kim J, Murphy TL, Murphy KM, Modlin RL, Shuai 
            K
  TITLE     PIASx is a transcriptional co-repressor of signal transducer and 
            activator of transcription 4.
  JOURNAL   J Biol Chem. 2003 Jun 13. 278(24):21327-30.
  PUBMED    12716907
REFERENCE   2  (bases 1 to 6519)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6519
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     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"
     regulatory      607..616
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             616..639
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    2544..3166
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2743..2762)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(3190..3210)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3195..3524
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3375..3510
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3437..3456
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     complement(3562..3581)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3563..3581)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3595..3611)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3595..3611)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3608..3630)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3619..3635
                     /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(3643..3673)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3688..3709
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3826..3843)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3997..4584)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4077..4096)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4755..5615)
                     /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     5378..5397
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5616..5720)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5747..6202
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5834..5853)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6044..6065
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6328..6350
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6342..6359
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6343..6359
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
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 ccgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttacggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta caccaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaataa ccccgccccg ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctcgttta gtgaaccgtc
      601 agaattgcca ccatggacta caaggacgac gatgacaaga attctgcagg atccaccatg
      661 gcggatttcg aagagttgag gaatatggtt tctagtttta gggtttctga actacaagta
      721 ttactaggct ttgctggacg gaataaaagt ggacgcaagc atgacctcct gatgagggcg
      781 ctgcatttat tgaagagcgg ctgcagccct gcggttcaga ttaaaatccg agaattgtat
      841 agacgccgat atccacgaac tcttgaagga ctttctgatt tatccacaat caaatcatcg
      901 gttttcagtt tggatggtgg ctcatcacct gtagaacctg acttggccgt ggctggaatc
      961 cactcgttgc cttccacttc agttacacct cactcaccat cctctcctgt tggttctgtg
     1021 ctgcttcaag atactaagcc cacatttgag atgcagcagc catctccccc aattcctcct
     1081 gtccatcctg atgtgcagtt aaaaaatctg cccttttatg atgtccttga tgttctcatc
     1141 aagcccacga gtttagttca aagcagtatt cagcgatttc aagagaagtt ttttattttt
     1201 gctttgacac ctcaacaagt tagagagata tgcatatcca gggatttttt gccaggtggt
     1261 aggagagatt atacagtcca agttcagttg agactttgcc tggcagagac aagttgccct
     1321 caagaagata actatccaaa tagtctatgt ataaaagtaa atgggaagct atttcctttg
     1381 cctggctatg caccaccgcc taaaaatggg attgaacaga agcgccctgg acgccccttg
     1441 aatattacat ctttagttag gttatcttca gctgtgccaa accaaatttc catttcttgg
     1501 gcatcagaaa ttgggaagaa ttactctatg tctgtatatc ttgtacggca gcttacatca
     1561 gccatgttat tacagagatt aaaaatgaaa ggtattagaa accctgatca ttccagagca
     1621 ctaattaaag aaaaacttac tgcagatcct gatagtgaaa ttgctacaac tagccttcgg
     1681 gtatccttga tgtgcccttt aggaaaaatg aggctgacaa tcccatgccg tgcagtgact
     1741 tgtacacatc tgcagtgttt tgatgctgcc ctctatctac aaatgaatga gaaaaagccc
     1801 acctggattt gtcctgtgtg tgacaaaaaa gctgcctatg aaagtctaat attagatggg
     1861 ctttttatgg aaattctcaa tgactgttct gatgtagatg agatcaaatt ccaagaagat
     1921 ggttcttggt gtccaatgag accgaagaaa gaagctatga aagtatccag ccaaccgtgt
     1981 acaaaaatag aaagttcaag cgtcctcagt aagccttgtt cagtgactgt agccagtgag
     2041 gcaagcaaga agaaagtaga tgttattgat cttacaatag aaagctcttc tgacgaagag
     2101 gaagaccctc ctgccaaaag gaaatgcatc tttatgtcag aaacacaaag cagcccaacc
     2161 aaaggggttc tcatgtatca gccatcttct gtaagggtgc ccagtgtgac ttcggttgat
     2221 cctgctgcta ttccgccttc attaacagac tactcagtac cattccacca tacgccaata
     2281 tcaagcatgt catcagattt gccaggtttg gattttcttt cccttattcc agttgatccc
     2341 cagtactgtc ctcctatgtt tttggatagt ctcacctcac ccttaacagc aagcagtacg
     2401 tctgtcacca ccaccagctc ccatgaaagc agtactcatg ttagttcatc cagcagcagg
     2461 agtgagacag gggtcataac cagcagtgga agtaacattc ctgaaatcat ctcattggac
     2521 taactcgaca tcgattaaag cttgggtggc atccctgtga cccctcccca gtgcctctcc
     2581 tggccctgga agttgccact ccagtgccca ccagccttgt cctaataaaa ttaagttgca
     2641 tcattttgtc tgactaggtg tccttctata atattatggg gtggaggggg gtggtatgga
     2701 gcaaggggca agttgggaag acaacctgta gggcctgcgg ggtctattgg gaaccaagct
     2761 ggagtgcagt ggcacaatct tggctcactg caatctccgc ctcctgggtt caagcgattc
     2821 tcctgcctca gcctcccgag ttgttgggat tccaggcatg catgaccagg ctcagctaat
     2881 ttttgttttt ttggtagaga cggggtttca ccatattggc caggctggtc tccaactcct
     2941 aatctcaggt gatctaccca ccttggcctc ccaaattgct gggattacag gcgtgaacca
     3001 ctgctccctt ccctgtcctt ctgattttaa aataactata ccagcaggag gacgtccaga
     3061 cacagcatag gctacctggc catgcccaac cggtgggaca tttgagttgc ttgcttggca
     3121 ctgtcctctc atgcgttggg tccactcagt agatgcctgt tgaattgggt acgcggccag
     3181 cttggctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca
     3241 gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct
     3301 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc
     3361 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
     3421 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc
     3481 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc ctcgaggaac
     3541 tgaaaaacca gaaagttaat tccctatagt gagtcgtatt aaattcgtaa tcatgtcata
     3601 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     3661 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     3721 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     3781 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     3841 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     3901 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     3961 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     4021 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     4081 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     4141 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     4201 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     4261 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     4321 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     4381 tgtaggcggt gctacagagt tcttgaagtg tggcctaact acggctacac tagaagaaca
     4441 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     4501 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     4561 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     4621 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     4681 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     4741 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     4801 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     4861 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     4921 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     4981 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     5041 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     5101 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     5161 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     5221 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     5281 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     5341 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     5401 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     5461 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     5521 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     5581 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     5641 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     5701 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgc gccctgtagc
     5761 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     5821 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     5881 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     5941 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag
     6001 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     6061 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg
     6121 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac
     6181 aaaatattaa cgcttacaat ttccattcgc cattcaggct gcgcaactgt tgggaagggc
     6241 gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc
     6301 gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggccagtg
     6361 ccaagctgat ctatacattg aatcaatatt ggcaattagc catattagtc attggttata
     6421 tagcataaat caatattggc tattggccat tgcatacgtt gtatctatat cataatatgt
     6481 acatttatat tggctcatgt ccaatatgac cgccatgtt
//
