LOCUS       sequence-169365-        8116 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8116)
  AUTHORS   Giancotti FG, Ruoslahti E
  TITLE     Elevated levels of the alpha 5 beta 1 fibronectin receptor suppress 
            the transformed phenotype of Chinese hamster ovary cells.
  JOURNAL   Cell. 1990 Mar 9. 60(5):849-59.
  PUBMED    2155708
REFERENCE   2  (bases 1 to 8116)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Cell. 1990 
            Mar 9. 60(5):849-59."
COMMENT     Sequence Label: pRK5 beta1
FEATURES             Location/Qualifiers
     source          1..8116
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    107..128
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(143..148,149..166,167..173)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    181..197
                     /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     186..208
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     205..221
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     205..221
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        250..629
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        630..833
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     784..804
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     830..854
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        1064..1082
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1064..1081
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     1095..1114
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      4652..4661
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    4665..4799
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4702..4721)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4756..4775
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        4868..5225
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4868..5059
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4891..4911)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5076..5211
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(5245..5262)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5245..5261)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5254..5276)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5474..5929
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5561..5580)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5771..5792
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5946..5965)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6065..6087
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6125..6143)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6211..6315
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6316..6384,6385..7176)
                     /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(6534..6553)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7347..7935
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7836..7855
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8089..8106
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
       61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
      121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
      181 ttgtgagcgg ataacaattt cacacaggaa acagctatga catgattacg aattaattcg
      241 agctcgcccg acattgatta ttgactagtt attaatagta atcaattacg gggtcattag
      301 ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct
      361 gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc
      421 caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg
      481 cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat
      541 ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
      601 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc
      661 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga
      721 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat
      781 tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctcgtttag
      841 tgaaccgtca gatcgcctgg agacgccatc cacgctgttt tgacctccat agaagacacc
      901 gggaccgatc cagcctccgc ggccgggaac ggtgcattgg aacgcggatt ccccgtgcca
      961 agagtgacgt aagtaccgcc tatagagtct ataggcccac ccccttggct tcgttagaac
     1021 gcggctacaa ttaatacata accttatgta tcatacacat acgatttagg tgacactata
     1081 gaataacatc cactttgcct ttctctccac aggtgtccac tcccaggtcc aactgcacct
     1141 cggttctatc gattgaattc cactcccgct gggccccgga cgccgcgcgg aaaagatgaa
     1201 tttacaacca attttctgga ttggactgat cagttcagtt tgcggtgtgt ttgctcaaac
     1261 agatgaaaat agatgtttaa aagcaaatgc caaatcatgt ggagaatgta tacaagcagg
     1321 gccaaattgt gggtggtgca caaattcaac atttttacag gaaggaatgc ctacttctgc
     1381 acgatgtgat gatttagaag ccttaaaaaa gaagggttgc cctccagatg acatagaaaa
     1441 tcccagaggc tccaaagata taaagaaaaa taaaaatgta accaaccgta gcaaaggaac
     1501 agcagagaag ctcaagccag aggatattac tcagatccaa ccacagcagt tggttttgcg
     1561 attaagatca ggggagccac agacatttac attaaaattc aagagagctg aagactatcc
     1621 cattgacctc tactacctta tggacctgtc ttattcaatg aaagacgatt tggagaatgt
     1681 aaaaagtctt ggaacagatc tgacgaatga aatgaggagg attacttcgg acttcagaat
     1741 tggatttggc tcatttgtgg aaaagactgt gatgccttac attagcacaa caccagctaa
     1801 gctcaggaac ccttgcacaa gtgaacagaa ctgcaccagc ccatttagct acaaaaatgt
     1861 gctcagtctt actagtaaag gagaagtatt taatgaactt gttggaaaac agcgcatatc
     1921 tggaaatttg gattctccag aaggtggttt cgatgccatc atgcaagttg cagtttgtgg
     1981 atcactgatt ggctggagga atgttacacg gctgctggtg ttttccacag atgccgggtt
     2041 tcactttgct ggagatggga aacttggtgg cattgtttta ccaaatgatg gacaatgtca
     2101 cctggaaaat aatatgtaca caatgagcca ttattatgat tatccttcta ttgctcacct
     2161 tgtccagaaa ctgagtgaaa ataatattca gacaattttt gcagttactg aagaatttca
     2221 gcctgtttac aaggagctga aaaacttgat ccctaagtca gcagtaggaa cattatctgc
     2281 caattctagc aatgtaattc agttgatcat tgatgcatac aattcccttt cctcagaagt
     2341 cattttggaa aacggcaaat tgtcagaagg cgtaacaata agttacaaat cttactgcaa
     2401 gaacggggtg aatggaacag gggaaaatgg aagaaaatgt tccaatattt ccattggaga
     2461 tgaggttcaa tttgaaatta gcataacttc aaataagtgt ccaaaaaagg attctgacag
     2521 ctttaaaatt aggcctctgg gctttacgga ggaagtagag gttattcttc agtacatctg
     2581 tgaatgtgaa tgccaaagcg aaggcatccc tgaaagtccc aagtgtcatg aaggaaatgg
     2641 gacatttgag tgtggcgcgt gcaggtgcaa tgaagggcgt gttggtagac attgtgaatg
     2701 cagcacagat gaagttaaca gtgaagacat ggatgcttac tgcaggaaag aaaacagttc
     2761 agaaatctgc agtaacaatg gagagtgcgt ctgcggacag tgtgtttgta ggaagaggga
     2821 taatacaaat gaaatttatt ctggcaaatt ctgcgagtgt gataatttca actgtgatag
     2881 atccaatggc ttaatttgtg gaggaaatgg tgtttgcaag tgtcgtgtgt gtgagtgcaa
     2941 ccccaactac actggcagtg catgtgactg ttctttggat actagtactt gtgaagccag
     3001 caacggacag atctgcaatg gccggggcat ctgcgagtgt ggtgtctgta agtgtacaga
     3061 tccgaagttt caagggcaaa cgtgtgagat gtgtcagacc tgccttggtg tctgtgctga
     3121 gcataaagaa tgtgttcagt gcagagcctt caataaagga gaaaagaaag acacatgcac
     3181 acaggaatgt tcctatttta acattaccaa ggtagaaagt cgggacaaat taccccagcc
     3241 ggtccaacct gatcctgtgt cccattgtaa ggagaaggat gttgacgact gttggttcta
     3301 ttttacgtat tcagtgaatg ggaacaacga ggtcatggtt catgttgtgg agaatccaga
     3361 gtgtcccact ggtccagaca tcattccaat tgtagctggt gtggttgctg gaattgttct
     3421 tattggcctt gcattactgc tgatatggaa gcttttaatg ataattcatg acagaaggga
     3481 gtttgctaaa tttgaaaagg agaaaatgaa tgccaaatgg gacacgggtg aaaatcctat
     3541 ttataagagt gccgtaacaa ctgtggtcaa tccgaagtat gagggaaaat gagtactgcc
     3601 cgtgcaaatc ccacaacact gaatgcaaag tagcaatttc catagtcaca gttaggtagc
     3661 tttagggcaa tattgccatg gttttactca tgtgcaggtt ttgaaaatgt acaatatgta
     3721 taatttttaa aatgttttat tattttgaaa ataatgttgt aattcatgcc agggactgac
     3781 aaaagacttg agacaggatg gttactcttg tcagctaagg tcacattgtg cctttttgac
     3841 cttttcttcc tggactattg aaatcaagct tattggatta agtgatattt ctatagcgat
     3901 tgaaagggca atagttaaag taatgagcat gatgagagtt tctgttaatc atgtattaaa
     3961 actgattttt agctttacaa atatgtcagt ttgcagttat gcagaatcca aagtaaatgt
     4021 cctgctagct agttaaggat tgttttaaat ctgttatttt gctatttgcc tgttagacat
     4081 gactgatgac atatctgaaa gacaagtatg ttgagagttg ctggtgtaaa atacgtttga
     4141 aatagttgat ctacaaaggc catgggaaaa attcagagag ttaggaagga aaaaccaata
     4201 gctttaaaac ctgtgtgcca ttttaagagt tacttaatgt ttggtaactt ttatgccttc
     4261 actttacaaa ttcaagcctt agataaaaga accgagcaat tttctgctaa aaagtccttg
     4321 atttagcact atttacatac aggccatact ttacaaagta tttgctgaat ggggaccttt
     4381 tgagttgaat ttattttatt atttttattt tgtttaatgt ctggtgcttt ctgtcacctc
     4441 ttctaatctt ttaatgtatt tgtttgcaat tttggggtaa gacttttttt atgagtactt
     4501 tttctttgaa gttttagcgg tcaatttgcc tttttaatga acatgtgaag ttatactgtg
     4561 gctatgcaac agctctcacc tacgcgagtc ttactttgag ttagtgccgg aattccccgg
     4621 ggatcctcta gagtcgacct gcagaagctt ggccgccatg gcccaacttg tttattgcag
     4681 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     4741 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctggatcg
     4801 ggaattaatt cggcgcagca ccatggcctg aaataacctc tgaaagagga acttggttag
     4861 gtaccttctg aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt
     4921 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     4981 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     5041 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
     5101 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     5161 cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
     5221 gcaaaaagct gttaacagct tggcactggc cgtcgtttta caacgtcgtg actgggaaaa
     5281 ccctggcgtt acccaactta atcgccttgc agcacatccc cctttcgcca gctggcgtaa
     5341 tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg
     5401 gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc gcatacgtca
     5461 aagcaaccat agtacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     5521 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct
     5581 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
     5641 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgattt gggtgatggt
     5701 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
     5761 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcgggctat
     5821 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt
     5881 taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt ttacaatttt atggtgcact
     5941 ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc
     6001 gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc
     6061 gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgagacga
     6121 aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag
     6181 acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
     6241 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
     6301 tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
     6361 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
     6421 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
     6481 gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt
     6541 ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat
     6601 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg
     6661 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
     6721 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
     6781 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
     6841 cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa
     6901 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca
     6961 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
     7021 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
     7081 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
     7141 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat
     7201 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     7261 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     7321 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     7381 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     7441 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
     7501 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     7561 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     7621 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     7681 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     7741 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     7801 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     7861 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     7921 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     7981 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
     8041 gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg
     8101 agcgaggaag cggaag
//
