LOCUS       Exported                7959 bp ds-DNA     circular SYN 28-MAY-2017
DEFINITION  pcDNA3.1/Puro-CAG-ASAP1 sequence 7389 bps.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7959)
  AUTHORS   Eric Schreiter
  TITLE     Direct Submission
  JOURNAL   Exported Oct 15, 2018 from SnapGene Viewer 4.2.6
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7959
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     CDS             14..1867
                     /codon_start=1
                     /label=VOLTRON_SOM2
                     /translation="MADVETETGMIAQWIVFAIMAAAAIAFGVAVHFRPSELKSAYYIN
                     IAICTIAATAYYAMAVNYQDLTMNGERQVVYARYINWVLTTPLLLLDLIVMTKMGGVMI
                     SWVIGADIFMIVFGILGAFEDEHKFKWVYFIAGCVMQAVLTYGMYNATWKDDLKKSPEY
                     HSSYVSLLVFLSILWVFYPVVWAFGSGSGVLSVDNEAILMGILDVLAKPLFGMGCLIAH
                     ETIFKIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSSYVWRNIIPHVAP
                     THRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVIHDWGSALGFHWA
                     KRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLIIDQNVFIEGTLPM
                     GVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALVEEYMDWLHQSPV
                     PKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDLIGSEIARWLSTL
                     EISGEPTTKSRITSEGEYIPLDQIDINVFCYENEVQSQPILNTKEMAPQSKPPEELEMS
                     SMPSPVAPLPARTEGVIDMRSMSSIDSFISCATDFPEATRF"
     CDS             14..694
                     /codon_start=1
                     /label=Ace2N
                     /translation="MADVETETGMIAQWIVFAIMAAAAIAFGVAVHFRPSELKSAYYIN
                     IAICTIAATAYYAMAVNYQDLTMNGERQVVYARYINWVLTTPLLLLDLIVMTKMGGVMI
                     SWVIGADIFMIVFGILGAFEDEHKFKWVYFIAGCVMQAVLTYGMYNATWKDDLKKSPEY
                     HSSYVSLLVFLSILWVFYPVVWAFGSGSGVLSVDNEAILMGILDVLAKPLFGMGCLIAH
                     ETIFK"
     CDS             695..1588
                     /codon_start=1
                     /label=HaloTag
                     /translation="IGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSSYVW
                     RNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVIHDW
                     GSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLIIDQN
                     VFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALVEEY
                     MDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDLIGS
                     EIARWLSTLEISGEPTT"
     CDS             1589..1648
                     /codon_start=1
                     /label=Kir2.1_membrane_trafficking_signal
                     /translation="KSRITSEGEYIPLDQIDINV"
     CDS             1649..1669
                     /codon_start=1
                     /label=Kir2.1_ER_export_signal
                     /translation="FCYENEV"
     CDS             1670..1864
                     /codon_start=1
                     /label=Kir2.1_proximal restriction and clustering signal
                     /translation="QSQPILNTKEMAPQSKPPEELEMSSMPSPVAPLPARTEGVIDMRS
                     MSSIDSFISCATDFPEATRF"
     polyA_signal    1988..2212
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2258..2686
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        2700..3029
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2880..3015
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3038..3637
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     polyA_signal    3797..3918
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3967..3983)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    3991..4007
                     /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(4015..4045)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4060..4081
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(4369..4954)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5125..5985)
                     /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"
     promoter        complement(5986..6090)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        6136..6515
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          6791..7807
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
ORIGIN
        1 gctagccgcc accatggctg acgtggaaac cgagaccggc atgattgcac agtggattgt
       61 ctttgctatt atggctgctg ctgctattgc ttttggagtg gctgtgcact ttcggccttc
      121 agagctgaag agcgcatact atatcaacat tgccatctgc actatcgccg ctaccgctta
      181 ctatgcaatg gccgtgaact accaggacct gacaatgaat ggtgaaaggc aggtggtcta
      241 cgcaagatat attaactggg tgctgaccac accactgctc ctgctcgatc tcatcgtcat
      301 gaccaagatg ggcggagtga tgatttcttg ggtcatcggc gcagacattt tcatgatcgt
      361 gtttggtatt ctgggcgcct tcgaggatga acacaagttc aaatgggtgt actttatcgc
      421 tggatgtgtg atgcaggcag tcctgacata cgggatgtat aacgccactt ggaaagacga
      481 tctgaagaaa agccccgagt accatagctc ctatgtcagt ctgctcgtct tcctgtcaat
      541 cctctgggtg ttttatcctg tcgtgtgggc tttcgggtct ggtagtggcg tgctgtccgt
      601 cgacaatgag gccattctca tgggaatcct ggatgtgctc gctaagccac tgtttggaat
      661 ggggtgcctc attgcccatg agactatctt caagatcggt actggctttc cattcgaccc
      721 ccattatgtg gaagtcctgg gcgagcgcat gcactacgtc gatgttggtc cgcgcgatgg
      781 cacccctgtg ctgttcctgc acggtaaccc gacctcctcc tacgtgtggc gcaacatcat
      841 cccgcatgtt gcaccgaccc atcgctgcat tgctccagac ctgatcggta tgggcaaatc
      901 cgacaaacca gacctgggtt atttcttcga cgaccacgtc cgcttcatgg atgccttcat
      961 cgaagccctg ggtctggaag aggtcgtcct ggtcattcac gactggggct ccgctctggg
     1021 tttccactgg gccaagcgca atccagagcg cgtcaaaggt attgcattta tggagttcat
     1081 ccgccctatc ccgacctggg acgaatggcc agaatttgcc cgcgagacct tccaggcctt
     1141 ccgcaccacc gacgtcggcc gcaagctgat catcgatcag aacgttttta tcgagggtac
     1201 gctgccgatg ggtgtcgtcc gcccgctgac tgaagtcgag atggaccatt accgcgagcc
     1261 gttcctgaat cctgttgacc gcgagccact gtggcgcttc ccaaacgagc tgccaatcgc
     1321 cggtgagcca gcgaacatcg tcgcgctggt cgaagaatac atggactggc tgcaccagtc
     1381 ccctgtcccg aagctgctgt tctggggcac cccaggcgtt ctgatcccac cggccgaagc
     1441 cgctcgcctg gccaaaagcc tgcctaactg caaggctgtg gacatcggcc cgggtctgaa
     1501 tctgctgcaa gaagacaacc cggacctgat cggcagcgag atcgcgcgct ggctgtcgac
     1561 gctcgagatt tccggcgagc caaccactaa gagcaggatc accagcgagg gcgagtacat
     1621 ccccctggac cagatcgaca tcaacgtgtt ctgctacgag aacgaggtgc aaagtcagcc
     1681 tatcctgaac acaaaggaaa tggctccaca gtctaagcct cccgaagagc ttgagatgtc
     1741 cagtatgcca agtcccgtgg ctcccctccc tgccaggact gaaggagtga ttgacatgag
     1801 gagtatgtca tctattgata gcttcatctc ttgcgcaaca gatttccccg aggctactcg
     1861 attctaatga aagcttggta ccgagctcgg atccactagt ccagtgtggt ggaattctgc
     1921 agatatccag cacagtggcg gccgctcgag tctagagggc ccgtttaaac ccgctgatca
     1981 gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc
     2041 ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg
     2101 cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg
     2161 gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat ggcttctgag
     2221 gcggaaagaa ccagctgggg ctctaggggg tatccccacg cgccctgtag cggcgcatta
     2281 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
     2341 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     2401 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     2461 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     2521 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     2581 acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc
     2641 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt ctgtggaatg
     2701 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     2761 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa
     2821 gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca
     2881 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
     2941 ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag tagtgaggag
     3001 gcttttttgg aggcctaggc ttttgcaaaa agctcccatg accgagtaca agcccacggt
     3061 gcgcctcgcc acccgcgacg acgtcccccg ggccgtacgc accctcgccg ccgcgttcgc
     3121 cgactacccc gccacgcgcc acaccgtcga cccggaccgc cacatcgagc gggtcaccga
     3181 gctgcaagaa ctcttcctca cgcgcgtcgg gctcgacatc ggcaaggtgt gggtcgcgga
     3241 cgacggcgcc gcggtggcgg tctggaccac gccggagagc gtcgaagcgg gggcggtgtt
     3301 cgccgagatc ggcccgcgca tggccgagtt gagcggttcc cggctggccg cgcagcaaca
     3361 gatggaaggc ctcctggcgc cgcaccggcc caaggagccc gcgtggttcc tggccaccgt
     3421 cggcgtctcg cccgaccacc agggcaaggg tctgggcagc gccgtcgtgc tccccggagt
     3481 ggaggcggcc gagcgcgccg gggtgcccgc cttcctggag acctccgcgc cccgcaacct
     3541 ccccttctac gagcggctcg gcttcaccgt caccgccgac gtcgaggtgc ccgaaggacc
     3601 gcgcacctgg tgcatgaccc gcaagcccgg tgcctgattc gaaatgaccg accaagcgac
     3661 gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt
     3721 cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga
     3781 gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     3841 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     3901 actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta
     3961 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
     4021 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
     4081 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     4141 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     4201 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     4261 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     4321 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     4381 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     4441 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     4501 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     4561 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     4621 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     4681 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     4741 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     4801 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     4861 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggttttt ttgtttgcaa
     4921 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     4981 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     5041 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     5101 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     5161 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     5221 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     5281 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     5341 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     5401 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     5461 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     5521 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     5581 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     5641 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     5701 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     5761 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     5821 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     5881 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     5941 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     6001 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     6061 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt
     6121 cgacggatcg gggtcgacat tgattattga ctagttatta atagtaatca attacggggt
     6181 cattagttca tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc
     6241 ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag
     6301 taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc
     6361 acttggcagt acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg
     6421 gtaaatggcc cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc
     6481 agtacatcta cgtattagtc atcgctatta ccatggtcga ggtgagcccc acgttctgct
     6541 tcactctccc catctccccc ccctccccac ccccaatttt gtatttattt attttttaat
     6601 tattttgtgc agcgatgggg gcgggggggg ggggcgcgcg ccaggcgggg cggggcgggg
     6661 cgaggggcgg ggcggggcga ggcggagagg tgcggcggca gccaatcaga gcggcgcgct
     6721 ccgaaagttt ccttttatgg cgaggcggcg gcggcggcgg ccctataaaa agcgaagcgc
     6781 gcggcgggcg ggagtcgctg cgcgctgcct tcgccccgtg ccccgctccg ccgccgcctc
     6841 gcgccgcccg ccccggctct gactgaccgc gttactccca caggtgagcg ggcgggacgg
     6901 cccttctcct ccgggctgta attagcgctt ggtttaatga cggcttgttt cttttctgtg
     6961 gctgcgtgaa agccttgagg ggctccggga gggccctttg tgcgggggga gcggctcggg
     7021 gggtgcgtgc gtgtgtgtgt gcgtggggag cgccgcgtgc ggctccgcgc tgcccggcgg
     7081 ctgtgagcgc tgcgggcgcg gcgcggggct ttgtgcgctc cgcagtgtgc gcgaggggag
     7141 cgcggccggg ggcggtgccc cgcggtgcgg ggggggctgc gaggggaaca aaggctgcgt
     7201 gcggggtgtg tgcgtggggg ggtgagcagg gggtgtgggc gcgtcggtcg ggctgcaacc
     7261 ccccctgcac ccccctcccc gagttgctga gcacggcccg gcttcgggtg cggggctccg
     7321 tacggggcgt ggcgcggggc tcgccgtgcc gggcgggggg tggcggcagg tgggggtgcc
     7381 gggcggggcg gggccgcctc gggccgggga gggctcgggg gaggggcgcg gcggcccccg
     7441 gagcgccggc ggctgtcgag gcgcggcgag ccgcagccat tgccttttat ggtaatcgtg
     7501 cgagagggcg cagggacttc ctttgtccca aatctgtgcg gagccgaaat ctgggaggcg
     7561 ccgccgcacc ccctctagcg ggcgcggggc gaagcggtgc ggcgccggca ggaaggaaat
     7621 gggcggggag ggccttcgtg cgtcgccgcg ccgccgtccc cttctccctc tccagcctcg
     7681 gggctgtccg cggggggacg gctgccttcg ggggggacgg ggcagggcgg ggttcggctt
     7741 ctggcgtgtg accggcggct ctagagcctc tgctaaccat gttcatgcct tcttcttttt
     7801 cctacagctc ctgggcaacg tgctggttat tgtgctgtct catcattttg gcaaagaatt
     7861 cctcgaggaa ttcactcctc aggtgcaggc tgcctatcag aaggtggtgg ctggtgtggc
     7921 caatgccctg gctcacaaat accactgaga tctgcatga
//
