LOCUS       sequence-258966-        6337 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 6337)
  TITLE     Susan Amara Plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6337)
  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            "Unpublished"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: CMV-hEAAT2
FEATURES             Location/Qualifiers
     source          1..6337
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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"
     polyA_signal    2361..2983
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2560..2579)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(3007..3027)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3012..3341
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3192..3327
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3379..3398)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3380..3398)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3412..3428)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3412..3428)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3425..3447)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3436..3452
                     /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(3460..3466,3467..3484,3485..3490))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3505..3526
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3643..3660)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3814..4402)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3894..3913)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4573..5364,5365..5433))
                     /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     5196..5215
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5434..5538)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5565..6020
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5652..5671)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5862..5883
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6146..6168
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6160..6177
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6161..6177
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
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 ccgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttacggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta caccaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaataa ccccgccccg ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctcgttta gtgaaccgtc
      601 agaattcaga tctggtacca ccatggcatc tacggaaggt gccaacaata tgcccaagca
      661 ggtggaagtg cgaatgcacg acagtcatct tggctcagag gaacccaagc accggcacct
      721 gggcctgcgc ctgtgtgaca agctggggaa gaatctgctg ctcaccctga cggtgtttgg
      781 tgtcatcctg ggagcagtgt gtggagggct tcttcgcttg gcatctccca tccaccctga
      841 tgtggttatg ttaatagcct tcccagggga tatactcatg aggatgctaa aaatgctcat
      901 tctccctcta atcatctcca gcttaatcac agggttgtca ggcctggatg ctaaggctag
      961 tggccgcttg ggcacgagag ccatggtgta ttacatgtcc acgaccatca ttgctgcagt
     1021 actgggggtc attctggtct tggctatcca tccaggcaat cccaagctca agaagcagct
     1081 ggggcctggg aagaagaatg atgaagtgtc cagcctggat gccttcctgg accttattcg
     1141 aaatctcttc cctgaaaacc ttgtccaagc ctgctttcaa cagattcaaa cagtgacgaa
     1201 gaaagtcctg gttgcaccac cgccagacga ggaggccaac gcaaccagcg ctgttgtctc
     1261 tctgttgaac gagactgtga ctgaggtgcc ggaggagact aagatggtta tcaagaaggg
     1321 cctggagttc aaggatggga tgaacgtctt aggtctgata gggtttttca ttgcttttgg
     1381 catcgctatg gggaagatgg gagatcaggc caagctgatg gtggatttct tcaacatttt
     1441 gaatgagatt gtaatgaagt tagtgatcat gatcatgtgg tactctcccc tgggtatcgc
     1501 ctgcctgatc tgtggaaaga tcattgcaat caaggactta gaagtggttg ctaggcaact
     1561 ggggatgtac atggtaacag tgatcatagg cctcatcatc cacgggggca tctttctccc
     1621 cttgatttac tttgtagtga ccaggaaaaa ccccttctcc ttttttgctg gcattttcca
     1681 agcttggatc actgccctgg gcaccgcttc cagtgctgga actttgcctg tcacctttcg
     1741 ttgcctggaa gaaaatctgg ggattgataa gcgtgtgact agattcgtcc ttcctgttgg
     1801 agcaaccatt aacatggatg gtacagccct ttatgaagcg gtggccgcca tctttatagc
     1861 ccaaatgaat ggtgttgtcc tggatggagg acagattgtg actgtaagcc tcacagccac
     1921 cctggcaagc gtcggcgcgg ccagtatccc cagtgccggg ctggtcacca tgctcctcat
     1981 tctgacagcc gtgggcctgc caacagagga catcagcctg ctggtggctg tggactggct
     2041 gctggacagg atgagaactt cagtcaatgt tgtgggtgac tcttttgggg ctgggatagt
     2101 ctatcacctc tccaagtctg agctggatac cattgactcc cagcatcgag tgcatgaaga
     2161 tattgaaatg accaagactc aatccattta tgatgacatg aagaaccaca gggaaagcaa
     2221 ctctaatcaa tgtgtctatg ctgcacacaa ctctgtcata gtagatgaat gcaaggtaac
     2281 tctggcagcc aatggaaagt cagccgactg cagtgttgag gaagaacctt ggaaacgtga
     2341 gaaataatct agaggatccc gggtggcatc cctgtgaccc ctccccagtg cctctcctgg
     2401 ccctggaagt tgccactcca gtgcccacca gccttgtcct aataaaatta agttgcatca
     2461 ttttgtctga ctaggtgtcc ttctataata ttatggggtg gaggggggtg gtatggagca
     2521 aggggcaagt tgggaagaca acctgtaggg cctgcggggt ctattgggaa ccaagctgga
     2581 gtgcagtggc acaatcttgg ctcactgcaa tctccgcctc ctgggttcaa gcgattctcc
     2641 tgcctcagcc tcccgagttg ttgggattcc aggcatgcat gaccaggctc agctaatttt
     2701 tgtttttttg gtagagacgg ggtttcacca tattggccag gctggtctcc aactcctaat
     2761 ctcaggtgat ctacccacct tggcctccca aattgctggg attacaggcg tgaaccactg
     2821 ctcccttccc tgtccttctg attttaaaat aactatacca gcaggaggac gtccagacac
     2881 agcataggct acctggccat gcccaaccgg tgggacattt gagttgcttg cttggcactg
     2941 tcctctcatg cgttgggtcc actcagtaga tgcctgttga attgggtacg cggccagctt
     3001 ggctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa
     3061 gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc
     3121 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc
     3181 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct
     3241 gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga
     3301 agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcctc gaggaactga
     3361 aaaaccagaa agttaattcc ctatagtgag tcgtattaaa ttcgtaatca tgtcatagct
     3421 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     3481 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     3541 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     3601 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     3661 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     3721 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     3781 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     3841 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     3901 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     3961 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     4021 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     4081 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     4141 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     4201 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     4261 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     4321 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     4381 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     4441 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     4501 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     4561 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     4621 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     4681 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     4741 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     4801 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     4861 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     4921 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     4981 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     5041 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     5101 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     5161 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     5221 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     5281 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     5341 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     5401 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     5461 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     5521 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgcgc cctgtagcgg
     5581 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     5641 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     5701 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
     5761 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac
     5821 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     5881 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat
     5941 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa
     6001 aatattaacg cttacaattt ccattcgcca ttcaggctgc gcaactgttg ggaagggcga
     6061 tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga
     6121 ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc
     6181 aagctgatct atacattgaa tcaatattgg caattagcca tattagtcat tggttatata
     6241 gcataaatca atattggcta ttggccattg catacgttgt atctatatca taatatgtac
     6301 atttatattg gctcatgtcc aatatgaccg ccatgtt
//
