LOCUS       sequence-6809-ex        4708 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 4708)
  TITLE     pRK7
REFERENCE   2  (bases 1 to 4708)
  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"
COMMENT     Sequence Label: pRK7
FEATURES             Location/Qualifiers
     source          1..4708
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        14..393
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        394..597
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     548..568
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     594..618
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        828..846
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     828..845
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     859..878
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      961..970
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    973..1107
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1010..1029)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1064..1083
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        1180..1537
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        1180..1371
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(1203..1223)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      1388..1523
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1450..1469
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     complement(1557..1574)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1557..1573)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1566..1588)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      1786..2241
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1873..1892)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2083..2104
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2258..2277)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2412..2434
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2487..2505)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2573..2677
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2678..2746,2747..3538)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYM
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRDDAGQEQLGRRIHYSQNDLVEYS
                     PATEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2896..2915)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3709..4297
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4198..4217
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4451..4468
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        join(4621..4626,4627..4644,4645..4651)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4659..4675
                     /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     4664..4686
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4683..4699
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4683..4699
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 ttcgagctcg cccgacattg attattgact agttattaat agtaatcaat tacggggtca
       61 ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct
      121 ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta
      181 acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac
      241 ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt
      301 aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag
      361 tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat
      421 gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat
      481 gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc
      541 ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctcgt
      601 ttagtgaacc gtcagatcgc ctggagacgc catccacgct gttttgacct ccatagaaga
      661 caccgggacc gatccagcct ccgcggccgg gaacggtgca ttggaacgcg gattccccgt
      721 gccaagagtg acgtaagtac cgcctataga gtctataggc ccaccccctt ggcttcgtta
      781 gaacgcggct acaattaata cataacctta tgtatcatac acatacgatt taggtgacac
      841 tatagaataa catccacttt gcctttctct ccacaggtgt ccactcccag gtccaactgc
      901 acctcggttc taagcttctg caggtcgact ctagaggatc cccggggaat tcaatcgatg
      961 gccgccatgg ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
     1021 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     1081 atcaatgtat cttatcatgt ctggatcgat cgggaattaa ttcggcgcag caccatggcc
     1141 tgaaataacc tctgaaagag gaacttggtt aggtaccttc tgaggcggaa agaaccagct
     1201 gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     1261 gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc
     1321 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac
     1381 tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact
     1441 aatttttttt atttatgcag aggccgaggc cgcctcggcc tgtgagctat tccagaagta
     1501 gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctgttaacag cttggcactg
     1561 gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg ttacccaact taatcgcctt
     1621 gcagcacatc cccccttcgc cacctggcgt aatagcgaag aggcccgcac cgatcgccct
     1681 tcccaacagt tgcgtagcct gaatggcgaa tggcgcctga tgcggtattt tctccttacg
     1741 catctgtgcg gtatttcaca ccgcatacgt caaagcaacc atagtacgcg ccctgtacgg
     1801 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg
     1861 ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc
     1921 cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc
     1981 tcgaccccaa aaaacttgat ttgggtgatg gttcacgtag tgggccatcg ccctgataga
     2041 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa
     2101 ctggaacaac actcaaccct atctcgggct attcttttga tttataaggg attttgccga
     2161 tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca
     2221 aaatattaac gtttacaatt ttatggtgca ctctcagtac aatctgctct gatgccgcat
     2281 agttaagcca actccgctat cgctacgtga ctgggtcatg gctgcgcccc gacacccgcc
     2341 aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc
     2401 tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc
     2461 gaggcagtat tcttgaagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt
     2521 catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac
     2581 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     2641 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
     2701 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
     2761 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tggaactgga
     2821 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
     2881 cacttttaaa gttctgctat gtggcgcggt attatcccgt gatgacgccg ggcaagagca
     2941 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagccacaga
     3001 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
     3061 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
     3121 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     3181 tgaagccata ccaaacgacg agcgtgacac cacgatgcca gcagcaatgg caacaacgtt
     3241 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
     3301 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
     3361 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg
     3421 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
     3481 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
     3541 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     3601 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     3661 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     3721 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     3781 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     3841 gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     3901 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     3961 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     4021 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     4081 gagataccta cagcgtgagc attgagaaag cgccacgctt cccgaaggga gaaaggcgga
     4141 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     4201 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     4261 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     4321 acggttcctg gccttttggt ggccttttgc tcacatgttc tttcctgcgt tatcccctga
     4381 ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac
     4441 gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc
     4501 tctccccgcg cgttggccga ttcattaatc cagctggcac gacaggtttc ccgactggaa
     4561 agcgggcagt gagcgcaacg caattaatgt gagttacctc actcattagg caccccaggc
     4621 tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca
     4681 cacaggaaac agctatgacc atgattac
//
