LOCUS       Stat3 Flag pRc/CMV                7969 bp ds-DNA     circular SYN 06-AUG-2018
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    Stat3 Flag pRc/CMV
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7969)
  AUTHORS   Horvath CM, Wen Z, Darnell JE Jr
  TITLE     A STAT protein domain that determines DNA sequence recognition 
            suggests a novel DNA-binding domain.
  JOURNAL   Genes Dev 1995 Apr 15;9(8):984-94.
  PUBMED    7774815
REFERENCE   2  (bases 1 to 7969)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 6, 2018 from SnapGene Server 1.1.58
            //www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7969
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     103..120
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    237..258
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        273..303
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    311..327
                     /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)."
     primer_bind     316..338
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     335..351
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     335..351
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    382..516
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(406..425)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     460..479
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(690..1484)
                     /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(801..820)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     1411..1430
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(1551..1880)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1565..1700
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(1619..1638)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     1865..1885
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(1908..2421)
                     /direction=LEFT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2103..2124)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2315..2334
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    2467..2691
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     2680..2697
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     promoter        2717..2735
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     2717..2734
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             complement(2744..2767)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(5175..5191)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5175..5191)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(5281..5300)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5282..5300)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(5345..5548)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(5374..5394)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        5549..5928
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     6100..6119
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        6194..6298
                     /gene="bla"
                     /label=AmpR promoter
     CDS             6299..7159
                     /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(6517..6536)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7330..7918
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7819..7838
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt
       61 gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag
      121 gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa
      181 tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat
      241 gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg
      301 ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac
      361 gccaagctct cgaggtcgac ggatccagac atgataagat acattgatga gtttggacaa
      421 accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga tgctattgct
      481 ttatttgtaa ccattataag ctgcaataaa caagttgggg tgggcgaaga actccagcat
      541 gagatccccg cgctggagga tcatccagcc ggcgtcccgg aaaacgattc cgaagcccaa
      601 cctttcatag aaggcggcgg tggaatcgaa atctcgtgat ggcaggttgg gcgtcgcttg
      661 gtcggtcatt tcgaacccca gagtcccgct cagaagaact cgtcaagaag gcgatagaag
      721 gcgatgcgct gcgaatcggg agcggcgata ccgtaaagca cgaggaagcg gtcagcccat
      781 tcgccgccaa gctcttcagc aatatcacgg gtagccaacg ctatgtcctg atagcggtcc
      841 gccacaccca gccggccaca gtcgatgaat ccagaaaagc ggccattttc caccatgata
      901 ttcggcaagc aggcatcgcc atgggtcacg acgagatcct cgccgtcggg catgcgcgcc
      961 ttgagcctgg cgaacagttc ggctggcgcg agcccctgat gctcttcgtc cagatcatcc
     1021 tgatcgacaa gaccggcttc catccgagta cgtgctcgct cgatgcgatg tttcgcttgg
     1081 tggtcgaatg ggcaggtagc cggatcaagc gtatgcagcc gccgcattgc atcagccatg
     1141 atggatactt tctcggcagg agcaaggtga gatgacagga gatcctgccc cggcacttcg
     1201 cccaatagca gccagtccct tcccgcttca gtgacaacgt cgagcacagc tgcgcaagga
     1261 acgcccgtcg tggccagcca cgatagccgc gctgcctcgt cctgcagttc attcagggca
     1321 ccggacaggt cggtcttgac aaaaagaacc gggcgcccct gcgctgacag ccggaacacg
     1381 gcggcatcag agcagccgat tgtctgttgt gcccagtcat agccgaatag cctctccacc
     1441 caagcggccg gagaacctgc gtgcaatcca tcttgttcaa tcatgcgaaa cgatcctcat
     1501 cctgtctctt gatcagatcc gaaaatggat atccaagctc ccgggagctt tttgcaaaag
     1561 cctaggcctc caaaaaagcc tcctcactac ttctggaata gctcagaggc cgaggcggcc
     1621 tcggcctctg cataaataaa aaaaattagt cagccatggg gcggagaatg ggcggaactg
     1681 ggcggagtta ggggcgggat gggcggagtt aggggcggga ctatggttgc tgactaattg
     1741 agatgcatgc tttgcatact tctgcctgct ggggagcctg gggactttcc acacctggtt
     1801 gctgactaat tgagatgcat gctttgcata cttctgcctg ctggggagcc tggggacttt
     1861 ccacacccta actgacacac attccacaga attcgagctc ggtacccacc ccggttgata
     1921 atcagaaaag ccccaaaaac aggaagattg tataagcaaa tatttaaatt gtaagcgtta
     1981 atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg
     2041 ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg
     2101 ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa
     2161 aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg
     2221 ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt
     2281 gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg
     2341 ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta
     2401 atgcgccgct acagggcgcg tggggatccc ccctcgagcc ccagctggtt ctttccgcct
     2461 cagaagccat agagcccacc gcatccccag catgcctgct attgtcttcc caatcctccc
     2521 ccttgctgtc ctgccccacc ccacccccca gaatagaatg acacctactc agacaatgcg
     2581 atgcaatttc ctcattttat taggaaagga cagtgggagt ggcaccttcc agggtcaagg
     2641 aaggcacggg ggaggggcaa acaacagatg gctggcaact agaaggcaca gtcgaggctg
     2701 atcagcgagc tctagcattt aggtgacact atagaatagg gcccttgtca tcgtcgtcct
     2761 tgtagtctct agacatgggg gaggtagcac actccgaggt cagatccatg tcaaacgtga
     2821 gcgactcaaa ctgccctcct gctgagggct cagcaccttc accgttattt ccaaactgca
     2881 tcaatgaatc taaagtgcgg ggggacatcg gcaggtcaat ggtattgctg caggtcgttg
     2941 gtgtcacaca gatgaacttg gtcttcaggt acggggcagc actacctggg tcggcttcgg
     3001 ggtgctcctg gctctcgggc ctacagtact ttccaaatgc ctcctccttg ggaatgtcgg
     3061 ggtagaggta gacaagtgga gacaccagga tgttggtcgc atccatgatc ttatagccca
     3121 tgatgatttc agcaaatgac atgttgttca gctgctgctt ggtgtatggc tctacagact
     3181 ggatctgggt cttgccactg atgtcctttt ccacccaagt gaaagtgacc cctccttctt
     3241 tgctgctctc gctgaagcgc agtaggaagg tgcccggggg ctttgtgctt aggatggccc
     3301 gctcccgctc cttgctgatg aaacccatga tgtacccttc attccaaagg gccaagatat
     3361 actttttcac aaggtcgatg atattgtcta gccagaccca gaaggagaag cccttgccag
     3421 ccatgttttc tttgcagaat ttagcccatg tgatctgaca ccctgagtag ttcacaccag
     3481 gccctaggag cttctcagcc agcgttgtca gctgctcgat gctcagccct cgcttggtgg
     3541 tggacgagaa ctgccagctg agcacctcgg ccacttggtc ccaggttcca attggcggct
     3601 tagtgaagaa gttcacgttc ttggggttat tggtcagcat gttataccac aggattgatg
     3661 cccaagcatt tggcatctga cagatgttgg agatcaccac aactggcaag gagtgggtct
     3721 ctaggtcaat cttgaggcct tggtggtaca cctcagtctc gaaggtgatc aggtgcagct
     3781 cctcagtcac gatcaaggag gcatcacaat tggcacggcc tccattccca catctctgct
     3841 ccctaagggt caggtgcttg aactctgcag acaggctgcc gttgttagac tcctccatgt
     3901 tcatcacttt tgtgttcgtg cccagaatgt taaatttccg agaccctctg agggcagcaa
     3961 catccccaga gtctttatca atgcacactt taattttaag ctgataattc aactcaggaa
     4021 atttgaccag caacctgact ttcgtggtaa actggacacc agtcttgatg actaagggcc
     4081 ggtccgggtg catgggcatg cagggctgcc gctccaccac gaaggcactc ttcattaagt
     4141 ttctgaacag ctccacgatc ctctcctcca gcatgggccg gtgctgcacg atagggtcgc
     4201 ccttgtagga cactttctgc tgcagctcct ccagtttctt aatttgttgg cgggtctgaa
     4261 gttgagattc tgctaatgaa gttatccagt tttccagacg gtccaggcag atgttgggag
     4321 ggcctccgat gcacgcgatc tgctgccgcc tcttccagtc agccagctct tcatcagtca
     4381 gtgtcttctg cacgtactcc attgctgaca agagccccgc cagctcactc acaatgcttc
     4441 tccgcatctg gtccagggct gtgagcatct gttccagctg ctgcatcttc tgtctggtca
     4501 cagactggtt gtttccattc agatcctgca tgtctccttg gctcttgagg gttttgtagt
     4561 tgaaatcaaa gtcgtcctgg aggttctcca ccaccttcat tttctgttct agatcctgca
     4621 ctcgcttccg gacatcctga agatgctgct ccaacatctg ctgcttctct gtcactacgg
     4681 cggctgttgg gtggttggcc tggccccctt gctgggctgc cgtggctgcc gtctggagga
     4741 ggcgagactc ttcccacagg catcgggcca cgatccgggc aatttccatt ggcttctcaa
     4801 gatacctgct ctgcagaaac tgcttgattc ttcgaaggtt gtgctgatag aggacattgg
     4861 actcttgcag gaatcggcta tattgctggt caatttcacc caagagatta tgaaacacca
     4921 acgtggcatg tgactctttg ctggctgcat atgcccagtc ttgactctca atccaaggtg
     4981 ccaggaactg ccgcagctcc atggggaagc tgtcgctgta cagctggtgc agctgcttca
     5041 ggtagcgtgt gtccagctgc tgcagctggt tccactgagc catcctgctg cagtcagggg
     5101 tctcgactgt ctccggggct gaggcccgac tggccggcct gcgctggtcg cggcggaatt
     5161 cctgcagccc gggggatcca ctagttctag agcggccgcc agtgtgatgg atatctgcag
     5221 aattccagca cactggcggc cgttactagt ggatccgagc tcggtaccaa gcttgggtct
     5281 ccctatagtg agtcgtatta atttcgataa gccagtaagc agtgggttct ctagttagcc
     5341 agagagctct gcttatatag acctcccacc gtacacgcct accgcccatt tgcgtcaatg
     5401 gggcggagtt gttacgacat tttggaaagt cccgttgatt ttggtgccaa aacaaactcc
     5461 cattgacgtc aatggggtgg agacttggaa atccccgtga gtcaaaccgc tatccacgcc
     5521 cattgatgta ctgccaaaac cgcatcacca tggtaatagc gatgactaat acgtagatgt
     5581 actgccaagt aggaaagtcc cataaggtca tgtactgggc ataatgccag gcgggccatt
     5641 taccgtcatt gacgtcaata gggggcgtac ttggcatatg atacacttga tgtactgcca
     5701 agtgggcagt ttaccgtaaa tactccaccc attgacgtca atggaaagtc cctattggcg
     5761 ttactatggg aacatacgtc attattgacg tcaatgggcg ggggtcgttg ggcggtcagc
     5821 caggcgggcc atttaccgta agttatgtaa cgcggaactc catatatggg ctatgaacta
     5881 atgaccccgt aattgattac tattaataac tagtcaataa tcaatgtcaa cgcgtatatc
     5941 tggcccgtac atcgcgaagc agcgcaaaac gcctaaccct aagcagattc ttcatgcaat
     6001 tgtcggtcaa gccttgcctt gttgtagctt aaattttgct cgcgcactac tcagcgacct
     6061 ccaacacaca agcagggagc agatactggc ttaactatgc ggcatcagag cagattgtac
     6121 tgagagtgca ccatagggga tcgggagatc tcccgatccg tcgacgtcag gtggcacttt
     6181 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     6241 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
     6301 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
     6361 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
     6421 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
     6481 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
     6541 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
     6601 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
     6661 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
     6721 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
     6781 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
     6841 tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
     6901 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
     6961 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
     7021 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
     7081 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
     7141 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
     7201 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     7261 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     7321 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     7381 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     7441 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     7501 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     7561 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     7621 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     7681 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     7741 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     7801 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     7861 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     7921 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggcctttt
//