LOCUS       sequence-73127-e        4829 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pCS2+ vector with MCS replaced by a TA-cloning linker .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4829)
  TITLE     pME-TA
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4829)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pCS2+TA
FEATURES             Location/Qualifiers
     source          1..4829
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        35..53
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     35..52
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             join(106..108,109..111,112..825)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(106..108,109..111,112..822)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(151..172)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(412..431)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     759..780
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    878..1012
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(902..921)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     956..975
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1125..1145)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1125..1143)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1164..1180)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1164..1180)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1177..1199)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1188..1204
                     /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(join(1212..1218,1219..1236,1237..1242))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1257..1278
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1395..1412)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1566..2154)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1646..1665)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2325..3116,3117..3185))
                     /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     2948..2967
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3186..3290)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3316..3771)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3453..3474)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3665..3684
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3845..4829
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
ORIGIN
        1 cgccattctg cctggggacg tcggagcaag cttgatttag gtgacactat agaatacaag
       61 ctacttgttc tttttgcagg atccccaata cttgtatggg atatcatggt gagcaagggc
      121 gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc
      181 cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg
      241 aagttcatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctg
      301 acctacggcg tgcagtgctt cagccgctac cccgaccaca tgaagcagca cgacttcttc
      361 aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc
      421 aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag
      481 ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac
      541 tacaacagcc acaacgtcta tatcatggcc gacaagcaga agaacggcat caaggtgaac
      601 ttcaagatcc gccacaacat cgaggacggc agcgtgcagc tcgccgacca ctaccagcag
      661 aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagcacccag
      721 tccgccctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg
      781 accgccgccg ggatcactct cggcatggac gagctgtaca agtaaccata caagtattgg
      841 ctcgagcctc tagaactata gtgagtcgta ttacgtagat ccagacatga taagatacat
      901 tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat
      961 ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag ttaacaacaa
     1021 caattgcatt cattttatgt ttcaggttca gggggaggtg tgggaggttt tttaattcgc
     1081 ggccgcggcg ccaatgcatt gggcccggta cccagctttt gttcccttta gtgagggtta
     1141 attgcgcgct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc
     1201 acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga
     1261 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
     1321 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     1381 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     1441 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     1501 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     1561 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     1621 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     1681 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     1741 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     1801 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     1861 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     1921 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     1981 tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca
     2041 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     2101 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     2161 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     2221 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     2281 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     2341 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     2401 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     2461 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     2521 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     2581 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     2641 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     2701 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     2761 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     2821 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     2881 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     2941 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     3001 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     3061 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     3121 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     3181 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     3241 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     3301 cgaaaagtgc cacctaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt
     3361 gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa
     3421 aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa
     3481 agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac
     3541 gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga
     3601 accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa
     3661 aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc
     3721 tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg tcccattcgc
     3781 cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc
     3841 agtcgaccat agccaattca atatggcgta tatggactca tgccaattca atatggtgga
     3901 tctggacctg tgccaattca atatggcgta tatggactcg tgccaattca atatggtgga
     3961 tctggacccc agccaattca atatggcgga cttggcacca tgccaattca atatggcgga
     4021 cttggcactg tgccaactgg ggaggggtct acttggcacg gtgccaagtt tgaggagggg
     4081 tcttggccct gtgccaagtc cgccatattg aattggcatg gtgccaataa tggcggccat
     4141 attggctata tgccaggatc aatatatagg caatatccaa tatggcccta tgccaatatg
     4201 gctattggcc aggttcaata ctatgtattg gccctatgcc atatagtatt ccatatatgg
     4261 gttttcctat tgacgtagat agcccctccc aatgggcggt cccatatacc atatatgggg
     4321 cttcctaata ccgcccatag ccactccccc attgacgtca atggtctcta tatatggtct
     4381 ttcctattga cgtcatatgg gcggtcctat tgacgtatat ggcgcctccc ccattgacgt
     4441 caattacggt aaatggcccg cctggctcaa tgcccattga cgtcaatagg accacccacc
     4501 attgacgtca atgggatggc tcattgccca ttcatatccg ttctcacgcc ccctattgac
     4561 gtcaatgacg gtaaatggcc cacttggcag tacatcaata tctattaata gtaacttggc
     4621 aagtacatta ctattggaag gacgccaggg tacattggca gtactcccat tgacgtcaat
     4681 ggcggtaaat ggcccgcgat ggctgccaag tacatcccca ttgacgtcaa tggggagggg
     4741 caatgacgca aatgggcgtt ccattgacgt aaatgggcgg taggcgtgcc taatgggagg
     4801 tctatataag caatgctcgt ttagggaac
//
