LOCUS       pcDNA3.1_rSERT_C        7423 bp DNA     circular SYN 29-JUL-2022
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7423)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 29, 2022 from SnapGene 6.1.0
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7423
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
                     /note="color: #ffffff"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
                     /note="color: #ffffff; direction: RIGHT"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /note="color: #ffffff; direction: RIGHT"
     CDS             991..1023
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /note="color: #cc99b2"
                     /translation="MEQKLISEEDL"
     CDS             1027..2985
                     /codon_start=1
                     /label=rSERT_C109A_Y107C
                     /note="color: #993366"
                     /translation="METTPLNSQKVLSECKDREDCQENGVLQKGVPTTADRAEPSQISN
                     GYSAVPSTSAGDEASHSIPAATTTLVAEIRQGERETWGKKMDFLLSVIGYAVDLGNIWR
                     FPCIAYQNGGGAFLLPYTIMAIFGGIPLFYMELALGQYHRNGCISIWRKICPIFKGIGY
                     AICIIAFYIASYYNTIIAWALYYLISSLTDRLPWTSCTNSWNTGNCTNYFAQDNITWTL
                     HSTSPAEEFYLRHVLQIHQSKGLQDLGTISWQLTLCIVLIFTVIYFSIWKGVKTSGKVV
                     WVTATFPYIVLSVLLVRGATLPGAWRGVVFYLKPNWQKLLETGVWVDAAAQIFFSLGPG
                     FGVLLAFASYNKFNNNCYQDALVTSVVNCMTSFVSGFVIFTVLGYMAEMRNEDVSEVAK
                     DAGPSLLFITYAEAIANMPASTFFAIIFFLMLITLGLDSTFAGLEGVITAVLDEFPHIW
                     AKRREWFVLIVVITCVLGSLLTLTSGGAYVVTLLEEYATGPAVLTVALIEAVAVSWFYG
                     ITQFCSDVKEMLGFSPGWFWRICWVAISPLFLLFIICSFLMSPPQLRLFQYNYPHWSIV
                     LGYCIGMSSVICIPTYIIYRLISTPGTLKERIIKSITPETPTEIPCGDIRMNAVAAAHH
                     HHHHGTDYKDDDDKEF*"
     CDS             2926..2943
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /note="color: #cc99b2"
                     /translation="HHHHHH"
     CDS             2950..2973
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /note="color: #cc99b2"
                     /translation="DYKDDDDK"
     polyA_signal    3023..3247
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
                     /note="color: #a6acb3"
     rep_origin      3293..3721
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
                     /note="color: #ffff00"
     promoter        3735..4064
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
                     /note="color: #ffffff; direction: RIGHT"
     rep_origin      3915..4050
                     /label=SV40 ori
                     /note="SV40 origin of replication"
                     /note="color: #ffff00"
     CDS             4131..4925
                     /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))"
                     /note="color: #ccffcc"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    5099..5220
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
                     /note="color: #a6acb3"
     primer_bind     complement(5269..5285)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
                     /note="color: #a020f0; direction: LEFT"
     protein_bind    5293..5309
                     /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)."
                     /note="color: #31849b"
     promoter        complement(5317..5347)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
                     /note="This reverse directional feature has 3 segments:
                        1: 5317 .. 5323 / #ffffff / -10
                        2: 5324 .. 5341 / #ffffff
                        3: 5342 .. 5347 / #ffffff / -35"
     protein_bind    5362..5383
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
                     /note="color: #31849b"
     rep_origin      complement(5671..6256)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #ffff00"
     CDS             complement(6427..7287)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /note="This feature has 2 segments:
                        1: 6427 .. 7218 / #ccffcc
                        2: 7219 .. 7287 / #ccffcc / signal sequence
                     Cleavage site after base 7218"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(7288..7392)
                     /gene="bla"
                     /label=AmpR promoter
                     /note="color: #ffffff; direction: LEFT"
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaactt aagcttggta ccgagctcgg atccactagt ccagtgtggt ggaattctgc
      961 agatatccag cacagtggcg gccgctcgag atggaacaaa aactcatttc agaagaggat
     1021 ctgaacatgg agaccacacc cttgaattca cagaaagtgc tgtcagagtg taaggacaga
     1081 gaggactgtc aagaaaatgg tgttctacag aagggtgtcc ccaccacagc ggacagggca
     1141 gagcctagcc aaatatccaa tgggtactct gcagtcccca gcacaagtgc aggggacgaa
     1201 gcttcacact cgatcccagc tgccaccacc accctggtgg ctgagattcg ccaaggggag
     1261 cgggagacct ggggcaagaa gatggatttc ctcctgtccg tcattggcta tgccgtggac
     1321 ctgggcaaca tctggcggtt tccttgcata gcctaccaga atggcggagg ggccttcctc
     1381 ctcccttata ccatcatggc cattttcggg gggatcccgc tcttttacat ggagctcgca
     1441 ctgggccagt accaccgaaa cgggtgcatt tccatatgga ggaagatctg cccgattttc
     1501 aaaggcattg gttacgccat ctgcatcatc gccttttaca tcgcctccta ctacaacacc
     1561 atcatagcct gggcgctcta ctacctcatc tcctccctca cggaccggct gccctggacc
     1621 agctgcacga actcctggaa cactggcaac tgcaccaact acttcgccca ggacaacatc
     1681 acctggacgc tgcattccac gtcccccgct gaggagttct acttgcgcca tgtcctgcag
     1741 atccaccagt ctaagggact ccaggacctg ggcaccatca gctggcagct gactctctgc
     1801 atcgtgctca tcttcaccgt aatctacttt agcatctgga aaggcgtcaa aacatctggc
     1861 aaggtggtgt gggtgacagc caccttccca tacattgtcc tctctgtcct gctggtgagg
     1921 ggggccaccc ttcctggagc ctggagaggg gtcgtcttct acttgaaacc caactggcag
     1981 aaactcttgg agacaggggt gtgggtagat gccgccgctc agatcttctt ctctcttggc
     2041 ccgggctttg gggttctcct ggcttttgct agctacaaca agttcaacaa caactgttac
     2101 caagatgccc tggtgaccag tgtggtgaac tgcatgacaa gcttcgtctc tggcttcgtc
     2161 atcttcacgg tgcttggcta catggcggag atgaggaatg aagatgtgtc agaggtggcc
     2221 aaagacgcag gccccagcct cctcttcatc acgtatgcag aggcaatagc caacatgcca
     2281 gcatccacgt tctttgccat catcttcttc ctcatgttaa tcacgctggg attggacagc
     2341 acgttcgcag gcctggaagg tgtgatcaca gctgtgctgg atgagttccc tcacatctgg
     2401 gccaagcgca gggaatggtt cgtgctcatc gtggtcatca cgtgcgtctt gggatccctg
     2461 ctcacactga cgtcaggagg ggcatacgtg gtgactctgc tggaggagta tgccacgggg
     2521 ccagcagtgc tcaccgtggc cctcatcgag gccgtcgccg tgtcttggtt ctatggaatc
     2581 actcagttct gcagcgatgt gaaggagatg ctgggcttca gcccgggatg gttttggagg
     2641 atctgctggg tggccatcag ccctctgttt ctcctgttca tcatttgcag ttttctgatg
     2701 agcccacccc agctacggct tttccaatac aactatcccc actggagtat cgtcttgggc
     2761 tactgcatag ggatgtcgtc cgtcatctgc atccctacct atatcattta tcggctgatc
     2821 agcactccgg ggacacttaa ggagcgcatt attaaaagta tcactcctga aacacccaca
     2881 gaaatcccgt gtggggacat ccgcatgaat gctgtggcgg ccgcacacca tcaccatcac
     2941 catggtaccg actacaagga cgacgatgac aaggaattct aataatctag agggcccgtt
     3001 taaacccgct gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc
     3061 tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat
     3121 gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg
     3181 caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc
     3241 tctatggctt ctgaggcgga aagaaccagc tggggctcta gggggtatcc ccacgcgccc
     3301 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     3361 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     3421 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     3481 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc
     3541 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     3601 ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt
     3661 ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     3721 taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca
     3781 gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct
     3841 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc
     3901 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
     3961 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc
     4021 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc ccgggagctt
     4081 gtatatccat tttcggatct gatcaagaga caggatgagg atcgtttcgc atgattgaac
     4141 aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact
     4201 gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc
     4261 gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg
     4321 cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg
     4381 tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt
     4441 catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc
     4501 atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag
     4561 cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg
     4621 ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc
     4681 tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt
     4741 ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg
     4801 ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt
     4861 acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct
     4921 tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg
     4981 agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga
     5041 cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa
     5101 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     5161 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     5221 tcatgtctgt ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt
     5281 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     5341 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     5401 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     5461 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg
     5521 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     5581 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     5641 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     5701 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     5761 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     5821 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     5881 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     5941 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     6001 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     6061 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     6121 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     6181 cggcaaacaa accaccgctg gtagcggttt ttttgtttgc aagcagcaga ttacgcgcag
     6241 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     6301 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     6361 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     6421 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     6481 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     6541 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     6601 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     6661 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     6721 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     6781 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     6841 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     6901 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     6961 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     7021 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     7081 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     7141 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     7201 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     7261 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     7321 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     7381 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtc
//
