LOCUS       Exported                7863 bp ds-DNA     circular SYN 28-SEP-2023
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    CD20
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7863)
  AUTHORS   van den Heuvel S, Harlow E
  TITLE     Distinct roles for cyclin-dependent kinases in cell cycle control.
  JOURNAL   Science 1993 Dec 24;262(5142):2050-4.
  PUBMED    8266103
REFERENCE   2  (bases 1 to 7863)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Sep 28, 2023 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7863
                     /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"
     primer_bind     581..605
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          696..1268
                     /label=-beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     primer_bind     complement(2785..2804)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2850..2905
                     /label=-beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2904..2923)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3263..3279)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3263..3279)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3276..3298)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3287..3303
                     /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(3311..3341)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3356..3377
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3494..3511)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3665..4253)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3745..3764)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4424..5284)
                     /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     5047..5066
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5285..5389)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5457..5475
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5513..5535)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5635..5654
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5848..5870
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5862..5879
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5863..5879
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        6290..6435
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     CDS             6492..7286
                     /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(6546..6565)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7156..7175
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(7480..7499)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    7524..7571
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
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 acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctcgttta gtgaaccgtc
      601 agatcgcctg gagacgccat ccacgctgtt ttgacctcca tagaagacac cgggaccgat
      661 ccagcctccg gtcgaccgat cctgagaact tcagggtgag tttggggacc cttgattgtt
      721 ctttcttttt cgctattgta aaattcatgt tatatggagg gggcaaagtt ttcagggtgt
      781 tgtttagaat gggaagatgt cccttgtatc accatggacc ctcatgataa ttttgtttct
      841 ttcactttct actctgttga caaccattgt ctcctcttat tttcttttca ttttctgtaa
      901 ctttttcgtt aaactttagc ttgcatttgt aacgaatttt taaattcact tttgtttatt
      961 tgtcagattg taagtacttt ctctaatcac ttttttttca aggcaatcag ggtatattat
     1021 attgtacttc agcacagttt tagagaacaa ttgttataat taaatgataa ggtagaatat
     1081 ttctgcatat aaattctggc tggcgtggaa atattcttat tggtagaaac aactacaccc
     1141 tggtcatcat cctgcctttc tctttatggt tacaatgata tacactgttt gagatgagga
     1201 taaaatactc tgagtccaaa ccgggcccct ctgctaacca tgttcatgcc ttcttctctt
     1261 tcctacagct cctgggcaac gtgctggttg ttgtgctgtc tcatcatttt ggcaaagaat
     1321 tcctcgacgg atccactcga gatccattgt gctctaaaga aatcagccct tgagatttga
     1381 ggccttggag actcaggagt tttgagagca aaatgacaac acccagaaat tcagtaaatg
     1441 ggactttccc ggcagagcca atgaaaggcc ctattgctat gcaatctggt ccaaaaccac
     1501 tcttcaggag gatgtcttca ctggtgggcc ccacgcaaag cttcttcatg agggaatcta
     1561 agactttggg ggctgtccag attatgaatg ggctcttcca cattgccctg gggggtcttc
     1621 tgatgatccc agcagggatc tatgcaccca tctgtgtgac tgtgtggtac cctctctggg
     1681 gaggcattat gtatattatt tccggatcac tcctggcagc aacggagaaa aactccagga
     1741 agtgtttggt caaaggaaaa atgataatga attcattgag cctctttgct gccatttctg
     1801 gaatgattct ttcaatcatg gacatactta atattaaaat ttcccatttt ttaaaaatgg
     1861 agagtctgaa ttttattaga gctcacacac catatattaa catatacaac tgtgaaccag
     1921 ctaatccctc tgagaaaaac tccccatcta cccaatactg ttacagcata caatctctgt
     1981 tcttgggcat tttgtcagtg atgctgatct ttgccttctt ccaggaactt gtaatagctg
     2041 gcatcgttga gaatgaatgg aaaagaacgt gctccagacc caaatctaac atagttctcc
     2101 tgtcagcaga agaaaaaaaa gaacagacta ttgaaataaa agaagaagtg gttgggctaa
     2161 ctgaaacatc ttcccaacca aagaatgaag aagacattga aattattcca atccaagaag
     2221 aggaagaaga agaaacagag acgaactttc cagaacctcc ccaagatcag gaatcctcac
     2281 caatagaaaa tgacagctct ccttaagtga tttcttctgt tttctgtttc cttttttaaa
     2341 cattagtgtt catagcttcc aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa
     2401 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa
     2461 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa
     2521 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa
     2581 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa
     2641 aaaaaaaaaa aaaaaaaaaa aaaactttag agcacaatgg atctcgagtg gatccgtcga
     2701 ggaattcact cctcaggtgc aggctgccta tcagaaggtg gtggctggtg tggccaatgc
     2761 cctggctcac aaataccact gagatctttt tccctctgcc aaaaattatg gggacatcat
     2821 gaagcccctt gagcatctga cttctggcta ataaaggaaa tttattttca ttgcaatagt
     2881 gtgttggaat tttttgtgtc tctcactcgg aaggacatat gggagggcaa atcatttaaa
     2941 acatcagaat gagtatttgg tttagagttt ggcaacatat gcccatatgc tggctgccat
     3001 gaacaaaggt tggctataaa gaggtcatca gtatatgaaa cagccccctg ctgtccattc
     3061 cttattccat agaaaagcct tgacttgagg ttagattttt tttatatttt gttttgtgtt
     3121 atttttttct ttaacatccc taaaattttc cttacatgtt ttactagcca gatttttcct
     3181 cctctcctga ctactcccag tcatagctgt ccctcttctc ttatggagat ccctcgacgg
     3241 atcggccgca attcgtaatc atgtcatagc tgtttcctgt gtgaaattgt tatccgctca
     3301 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     3361 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     3421 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     3481 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     3541 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     3601 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     3661 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     3721 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     3781 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     3841 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     3901 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     3961 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     4021 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     4081 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     4141 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     4201 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     4261 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     4321 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     4381 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     4441 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     4501 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     4561 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     4621 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     4681 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     4741 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     4801 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     4861 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     4921 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     4981 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     5041 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     5101 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     5161 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     5221 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     5281 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     5341 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     5401 gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata
     5461 ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac
     5521 acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag
     5581 cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat
     5641 cagagcagat tgtactgaga gtgcaccata tgcggtgtga aataccgcac agatgcgtaa
     5701 ggagaaaata ccgcatcagg cgccattcgc cattcaggct gcgcaactgt tgggaagggc
     5761 gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc
     5821 gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggccagtg
     5881 ccaagcttca cgctgccgca agcactcagg gcgcaagggc tgctaaagga agcggaacac
     5941 gtagaaagcc agtccgcaga aacggtgctg accccggatg aatgtcagct actgggctat
     6001 ctggacaagg gaaaacgcaa gcgcaaagag aaagcaggta gcttgcagtg ggcttacatg
     6061 gcgatagcta gactgggcgg ttttatggac agcaagcgaa ccggaattgc cagctggggc
     6121 gccctctggt aaggttggga agccctgcaa agtaaactgg atggctttct tgccgccaag
     6181 gatctgatgg cgcaggggat caagatcctg cttcatcccc gtggcccgtt gctcgcgttt
     6241 gctggcggtg tccccggaag aaatatattt gcatgtcttt agttctatga tgacacaaac
     6301 cccgcccagc gtcttgtcat tggcgaattc gaacacgcag atgcagtcgg ggcggcgcgg
     6361 tcccaggtcc acttcgcata ttaaggtgac gcgtgtggcc tcgaacaccg agcgaccctg
     6421 cagcgacccg cttaacagcg tcaacagcgt gccgcagatc tgatcaagag acaggatgag
     6481 gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg
     6541 agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt
     6601 tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc
     6661 tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt
     6721 gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag
     6781 tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg
     6841 ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag
     6901 cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg
     6961 atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc
     7021 gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca
     7081 tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc
     7141 gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg
     7201 ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct
     7261 atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc
     7321 gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg
     7381 cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct
     7441 ggagttcttc gcccaccccg ggagatgggg gaggctaact gaaacacgga aggagacaat
     7501 accggaagga acccgcgcta tgacggcaat aaaaagacag aataaaacgc acggtgttgg
     7561 gtcgtttgtt cataaacgcg gggttcggtc ccagggctgg cactctgtcg ataccccacc
     7621 gagaccccat tggggccaat acgcccgcgt ttcttccttt tccccacccc accccccaag
     7681 ttcgggtgaa ggcccagggc tcgcagccaa cgtcggggcg gcaagccctg ccatagccac
     7741 gggccccgtg ggttagggac ggcggatcgc ggccctctag agagcttggc ccattgcata
     7801 cgttgtatcc atatcataat atgtacattt atattggctc atgtccaaca ttaccgccat
     7861 gtt
//
