LOCUS       Exported                7481 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pMYC-GFP
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7481)
  AUTHORS   Chen Y, Xu J, Borowicz S, Collins C, Huo D, Olopade OI
  TITLE     c-Myc activates BRCA1 gene expression through distal promoter 
            elements in breast cancer cells.
  JOURNAL   BMC Cancer. 2011 Jun 13;11:246. doi: 10.1186/1471-2407-11-246.
  PUBMED    21668996
REFERENCE   2  (bases 1 to 7481)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7481
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     protein_bind    55..76
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        91..121
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    129..145
                     /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     134..156
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     153..169
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     153..169
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    218..339
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(229..248)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     283..302
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(515..1309)
                     /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(626..645)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     1236..1255
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(1376..1705)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1390..1525
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(1444..1463)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     1690..1710
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(1719..2147)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1829..1850)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2041..2060
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    2193..2417
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     2406..2423
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(2445..3164)
                     /codon_start=1
                     /product="Cycle 3 GFP (Crameri et al., 1996)"
                     /label=Cycle 3 GFP
                     /note="mammalian codon-optimized"
                     /translation="MASKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFSYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     primer_bind     complement(2488..2508)
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     3103..3131
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     CDS             complement(3216..4529)
                     /codon_start=1
                     /product="human c-Myc proto-oncogene"
                     /label=c-Myc
                     /translation="MPLNVSFTNRNYDLDYDSVQPYFYCDEEENFYQQQQQSELQPPAP
                     SEDIWKKFELLPTPPLSPSRRSGLCSPSYVAVTPFSLRGDNDGGGGSFSTADQLEMVTE
                     LLGGDMVNQSFICDPDDETFIKNIIIQDCMWSGFSAAAKLVSEKLASYQAARKDSGSPN
                     PARGHSVCSTSSLYLQDLSAAASECIDPSVVFPYPLNDSSSPKSCASQDSSAFSPSSDS
                     LLSSTESSPQGSPEPLVLHEETPPTTSSDSEEEQEDEEEIDVVSVEKRQAPGKRSESGS
                     PSAGGHSKPPHSPLVLKRCHVSTHQHNYAAPPSTRKDYPAAKRVKLDSVRVLRQISNNR
                     KCTSPRSSDTEENVKRRTHNVLERQRRNELKRSFFALRDQIPELENNEKAPKVVILKKA
                     TAYILSVQAEEQKLISEEDLLRKRREQLKHKLEQLRNSC"
     primer_bind     complement(4611..4630)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4612..4630)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(4675..4878)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(4704..4724)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        4879..5258
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     5430..5449
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        5524..5628
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5629..6489
                     /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(5847..5866)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6660..7248
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7149..7168
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7402..7419
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt
       61 gagttagctc actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt
      121 gtgtggaatt gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgc
      181 caagctctag ctagaggtcg acggtataca gacatgataa gatacattga tgagtttgga
      241 caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt
      301 gctttatttg taaccattat aagctgcaat aaacaagttg gggtgggcga agaactccag
      361 catgagatcc ccgcgctgga ggatcatcca gccggcgtcc cggaaaacga ttccgaagcc
      421 caacctttca tagaaggcgg cggtggaatc gaaatctcgt gatggcaggt tgggcgtcgc
      481 ttggtcggtc atttcgcgaa ccccagagtc ccgctcagaa gaactcgtca agaaggcgat
      541 agaaggcgat gcgctgcgaa tcgggagcgg cgataccgta aagcacgagg aagcggtcag
      601 cccattcgcc gccaagctct tcagcaatat cacgggtagc caacgctatg tcctgatagc
      661 ggtccgccac acccagccgg ccacagtcga tgaatccaga aaagcggcca ttttccacca
      721 tgatattcgg caagcaggca tcgccatggg tcacgacgag atcctcgccg tcgggcatgc
      781 gcgccttgag cctggcgaac agttcggctg gcgcgagccc ctgatgctct tcgtccagat
      841 catcctgatc gacaagaccg gcttccatcc gagtacgtgc tcgctcgatg cgatgtttcg
      901 cttggtggtc gaatgggcag gtagccggat caagcgtatg cagccgccgc attgcatcag
      961 ccatgatgga tactttctcg gcaggagcaa ggtgagatga caggagatcc tgccccggca
     1021 cttcgcccaa tagcagccag tcccttcccg cttcagtgac aacgtcgagc acagctgcgc
     1081 aaggaacgcc cgtcgtggcc agccacgata gccgcgctgc ctcgtcctgc agttcattca
     1141 gggcaccgga caggtcggtc ttgacaaaaa gaaccgggcg cccctgcgct gacagccgga
     1201 acacggcggc atcagagcag ccgattgtct gttgtgccca gtcatagccg aatagcctct
     1261 ccacccaagc ggccggagaa cctgcgtgca atccatcttg ttcaatcatg cgaaacgatc
     1321 ctcatcctgt ctcttgatca gatccgaaaa tggatataca agctcccggg agctttttgc
     1381 aaaagcctag gcctccaaaa aagcctcctc actacttctg gaatagctca gaggcagagg
     1441 cggcctcggc ctctgcataa ataaaaaaaa ttagtcagcc atggggcgga gaatgggcgg
     1501 aactgggcgg agttaggggc gggatgggcg gagttagggg cgggactatg gttgctgact
     1561 aattgagatg catgctttgc atacttctgc ctgctgggga gcctggggac tttccacacc
     1621 tggttgctga ctaattgaga tgcatgcttt gcatacttct gcctgctggg gagcctgggg
     1681 actttccaca ccctaactga cacacattcc acagaattaa ttcgcgttaa atttttgtta
     1741 aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga
     1801 atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa
     1861 cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga
     1921 accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc
     1981 taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga
     2041 agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg
     2101 cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtggg gataccccct
     2161 agagccccag ctggttcttt ccgcctcaga agccatagag cccaccgcat ccccagcatg
     2221 cctgctattg tcttcccaat cctccccctt gctgtcctgc cccaccccac cccccagaat
     2281 agaatgacac ctactcagac aatgcgatgc aatttcctca ttttattagg aaaggacagt
     2341 gggagtggca ccttccaggg tcaaggaagg cacgggggag gggcaaacaa cagatggctg
     2401 gcaactagaa ggcacagtcg aggctgatca gcgggtttaa ttcattattt gtagagctca
     2461 tccatgccat gtgtaatccc agcagcagtt acaaactcaa gaaggaccat gtggtcacgc
     2521 ttttcgttgg gatctttcga aagggcagat tgtgtcgaca ggtaatggtt gtctggtaaa
     2581 aggacagggc catcgccaat tggagtattt tgttgataat ggtctgctag ttgaacggat
     2641 ccatcttcaa tgttgtggcg aattttgaag ttagctttga ttccattctt ttgtttgtct
     2701 gccgtgatgt atacattgtg tgagttatag ttgtactcga gtttgtgtcc gagaatgttt
     2761 ccatcttctt taaaatcaat accttttaac tcgatacgat taacaagggt atcaccttca
     2821 aacttgactt cagcacgcgt cttgtagttc ccgtcatctt tgaaagatat agtgcgttcc
     2881 tgtacataac cttcgggcat ggcactcttg aaaaagtcat gccgtttcat atgatccgga
     2941 taacgggaaa agcattgaac accataagag aaagtagtga caagtgttgg ccatggaaca
     3001 ggtagttttc cagtagtgca aataaattta agggtaagct ttccgtatgt agcatcacct
     3061 tcaccctctc cactgacaga aaatttgtgc ccattaacat caccatctaa ttcaacaaga
     3121 attgggacaa ctccagtgaa aagttcttct cctttgctag ccattctaga ctcgagcggc
     3181 cgccactgtg ctggatatct gcagaattgc ccttcacaag agttccgtag ctgttcaagt
     3241 ttgtgtttca actgttctcg tcgtttccgc aacaagtcct cttcagaaat gagcttttgc
     3301 tcctctgctt ggacggacag gatgtatgct gtggcttttt taaggataac taccttgggg
     3361 gccttttcat tgttttccaa ctccgggatc tggtcacgca gggcaaaaaa gctccgtttt
     3421 agctcgttcc tcctctggcg ctccaagacg ttgtgtgttc gcctcttgac attctcctcg
     3481 gtgtccgagg acctggggct ggtgcatttt cggttgttgc tgatctgtct caggactctg
     3541 acactgtcca acttgaccct cttggcagca ggatagtcct tccgagtgga gggaggcgct
     3601 gcgtagttgt gctgatgtgt ggagacgtgg cacctcttga ggaccagtgg gctgtgagga
     3661 ggtttgctgt ggcctccagc agaaggtgat ccagactctg accttttgcc aggagcctgc
     3721 ctcttttcca cagaaacaac atcgatttct tcctcatctt cttgttcctc ctcagagtcg
     3781 ctgctggtgg tgggcggtgt ctcctcatgg agcaccaggg gctcggggct gccctgcggg
     3841 gaggactccg tcgaggagag cagagaatcc gaggacggag agaaggcgct ggagtcttgc
     3901 gaggcgcagg acttgggcga gctgctgtcg ttgagagggt aggggaagac caccgagggg
     3961 tcgatgcact ctgaggcggc ggcgctcaga tcctgcaggt acaagctgga ggtggagcag
     4021 acgctgtggc cgcgggcggg gttcgggctg ccgctgtctt tgcgcgcagc ctggtaggag
     4081 gccagcttct ctgagacgag cttggcggcg gccgagaagc cgctccacat acagtcctgg
     4141 atgatgatgt ttttgatgaa ggtctcgtcg tccgggtcgc agatgaaact ctggttcacc
     4201 atgtctcctc ccagcagctc ggtcaccatc tccagctggt cggccgtgga gaagctcccg
     4261 ccaccgccgt cgttgtctcc ccgaagggag aagggtgtga ccgcaacgta ggagggcgag
     4321 cagagcccgg agcggcggct aggggacagg ggcggggtgg gcagcagctc gaatttcttc
     4381 cagatatcct cgctgggcgc cgggggctgc agctcgctct gctgctgctg ctggtagaag
     4441 ttctcctcct cgtcgcagta gaaatacggc tgcaccgagt cgtagtcgag gtcatagttc
     4501 ctgttggtga agctaacgtt gaggggcatc gtcgcgggaa agggcaattc caccacactg
     4561 gactagtgga tccgagctcg gtaccaagct taactagcca gcttgggtct ccctatagtg
     4621 agtcgtatta atttcgataa gccagtaagc agtgggttct ctagttagcc agagagctct
     4681 gcttatatag acctcccacc gtacacgcct accgcccatt tgcgtcaatg gggcggagtt
     4741 gttacgacat tttggaaagt cccgttgatt ttggtgccaa aacaaactcc cattgacgtc
     4801 aatggggtgg agacttggaa atccccgtga gtcaaaccgc tatccacgcc cattgatgta
     4861 ctgccaaaac cgcatcacca tggtaatagc gatgactaat acgtagatgt actgccaagt
     4921 aggaaagtcc cataaggtca tgtactgggc ataatgccag gcgggccatt taccgtcatt
     4981 gacgtcaata gggggcgtac ttggcatatg atacacttga tgtactgcca agtgggcagt
     5041 ttaccgtaaa tactccaccc attgacgtca atggaaagtc cctattggcg ttactatggg
     5101 aacatacgtc attattgacg tcaatgggcg ggggtcgttg ggcggtcagc caggcgggcc
     5161 atttaccgta agttatgtaa cgcggaactc catatatggg ctatgaacta atgaccccgt
     5221 aattgattac tattaataac tagtcaataa tcaatgtcaa cgcgtatatc tggcccgtac
     5281 atcgcgaagc agcgcaaaac gcctaaccct aagcagattc ttcatgcaat tgtcggtcaa
     5341 gccttgcctt gttgtagctt aaattttgct cgcgcactac tcagcgacct ccaacacaca
     5401 agcagggagc agatactggc ttaactatgc ggcatcagag cagattgtac tgagagtgca
     5461 ccatagggga tcgggagatc tcccgatccg tcgacgtcag gtggcacttt tcggggaaat
     5521 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     5581 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa
     5641 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
     5701 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
     5761 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
     5821 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc
     5881 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
     5941 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
     6001 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
     6061 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
     6121 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg
     6181 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
     6241 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
     6301 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt
     6361 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
     6421 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
     6481 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat
     6541 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     6601 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     6661 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     6721 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     6781 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
     6841 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     6901 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     6961 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     7021 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     7081 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     7141 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     7201 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     7261 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     7321 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
     7381 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata
     7441 cgcaaaccgc ctctccccgc gcgttggccg attcattaat g
//
