LOCUS       sequence-456466-        6181 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 6181)
  TITLE     Susan Amara Plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6181)
  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            "Unpublished"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-hEAAT3
FEATURES             Location/Qualifiers
     source          1..6181
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     136..158
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     150..167
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     151..167
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        328..707
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        708..911
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     862..882
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     polyA_signal    2532..3154
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2731..2750)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(3178..3198)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3183..3512
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3363..3498
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3550..3569)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3551..3569)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3583..3599)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3583..3599)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3596..3618)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3607..3623
                     /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(3631..3637,3638..3655,3656..3661))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3676..3697
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3814..3831)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3985..4573)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4065..4084)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4744..5535,5536..5604))
                     /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     5367..5386
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5605..5709)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5736..6164
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5823..5842)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6033..6054
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 cttacaattt ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg
       61 cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg
      121 taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc aagctgatct
      181 atacattgaa tcaatattgg caattagcca tattagtcat tggttatata gcataaatca
      241 atattggcta ttggccattg catacgttgt atctatatca taatatgtac atttatattg
      301 gctcatgtcc aatatgaccg ccatgttgac attgattatt gactagttat taatagtaat
      361 caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg
      421 taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt
      481 atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac
      541 ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg
      601 acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact
      661 ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt
      721 ggcagtacac caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc
      781 ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc
      841 gtaataaccc cgccccgttg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata
      901 taagcagagc tcgtttagtg aaccgtcaga attcagatct ggtaccgcca tggggaaacc
      961 ggcgaggaaa ggatgcgagt ggaagcgctt cctgaagaat aactgggtgt tgctgtccac
     1021 cgtggccgcg gtggtgctag gcattaccac aggagtcttg gttcgagaac acagcaacct
     1081 ctcaactcta gagaaattct actttgcttt tcctggagaa attctaatgc ggatgctgaa
     1141 actcatcatt ttgccattaa ttatatccag catgattaca ggtgttgctg cactggattc
     1201 caacgtatcc ggaaaaattg gtctgcgcgc tgtcgtgtat tatttctgta ccactctcat
     1261 tgctgttatt ctaggtattg tgctggtggt gagcatcaag cctggtgtca cccagaaagt
     1321 gggtgaaatt gcgaggacag gcagcacccc tgaagtcagt acggtggatg ccatgttaga
     1381 tctcatcagg aatatgttcc ctgagaatct tgtccaggcc tgttttcagc agtacaaaac
     1441 taagcgtgaa gaagtgaagc ctcccagcga tccagagatg aacatgacag aagagtcctt
     1501 cacagctgtc atgacaactg caatttccaa gaacaaaaca aaggaataca aaattgttgg
     1561 catgtattca gatggcataa acgtcctggg cttgattgtc ttttgccttg tctttggact
     1621 tgtcattgga aaaatgggag aaaagggaca aattctggtg gatttcttca atgctttgag
     1681 tgatgcaacc atgaaaatcg ttcagatcat catgtgttat atgccactag gtattttgtt
     1741 cctgattgct gggaagatca tagaagttga agactgggaa atattccgca agctgggcct
     1801 ttacatggcc acagtcctga ctgggcttgc aatccactcc attgtaattc tcccgctgat
     1861 atatttcata gtcgtacgaa agaacccttt ccgatttgcc atgggaatgg cccaggctct
     1921 cctgacagct ctcatgatct cttccagttc agcaacactg cctgtcacct tccgctgtgc
     1981 tgaagaaaat aaccaggtgg acaagaggat cactcgattc gtgttacccg ttggtgcaac
     2041 aatcaacatg gatgggaccg cgctctatga agcagtggca gcggtgttta ttgcacagtt
     2101 gaatgacctg gacttgggca ttgggcagat catcaccatc agtatcacgg ccacatctgc
     2161 cagcatcgga gctgctggcg tgccccaggc tggcctggtg accatggtga ttgtgctgag
     2221 tgccgtgggc ctgcccgccg aggatgtcac cctgatcatt gctgtcgact ggctcctgga
     2281 ccggttcagg accatggtca acgtccttgg tgatgctttt gggacgggca ttgtggaaaa
     2341 gctctccaag aaggagctgg agcagatgga tgtttcatct gaagtcaaca ttgtgaatcc
     2401 ctttgccttg gaatccacaa tccttgacaa cgaagactca gacaccaaga agtcttatgt
     2461 caatggaggc tttgcagtag acaagtctga caccatctca ttcacccaga cctcacagtt
     2521 ctagggatcc cgggtggcat ccctgtgacc cctccccagt gcctctcctg gccctggaag
     2581 ttgccactcc agtgcccacc agccttgtcc taataaaatt aagttgcatc attttgtctg
     2641 actaggtgtc cttctataat attatggggt ggaggggggt ggtatggagc aaggggcaag
     2701 ttgggaagac aacctgtagg gcctgcgggg tctattggga accaagctgg agtgcagtgg
     2761 cacaatcttg gctcactgca atctccgcct cctgggttca agcgattctc ctgcctcagc
     2821 ctcccgagtt gttgggattc caggcatgca tgaccaggct cagctaattt ttgttttttt
     2881 ggtagagacg gggtttcacc atattggcca ggctggtctc caactcctaa tctcaggtga
     2941 tctacccacc ttggcctccc aaattgctgg gattacaggc gtgaaccact gctcccttcc
     3001 ctgtccttct gattttaaaa taactatacc agcaggagga cgtccagaca cagcataggc
     3061 tacctggcca tgcccaaccg gtgggacatt tgagttgctt gcttggcact gtcctctcat
     3121 gcgttgggtc cactcagtag atgcctgttg aattgggtac gcggccagct tggctgtgga
     3181 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     3241 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca
     3301 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
     3361 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     3421 tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag
     3481 gaggcttttt tggaggccta ggcttttgca aaaagctcct cgaggaactg aaaaaccaga
     3541 aagttaattc cctatagtga gtcgtattaa attcgtaatc atgtcatagc tgtttcctgt
     3601 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
     3661 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
     3721 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
     3781 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     3841 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     3901 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     3961 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     4021 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     4081 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     4141 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     4201 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     4261 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     4321 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     4381 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     4441 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     4501 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     4561 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     4621 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     4681 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     4741 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     4801 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     4861 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     4921 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     4981 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     5041 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     5101 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     5161 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     5221 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     5281 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     5341 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     5401 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     5461 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     5521 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     5581 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     5641 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     5701 ggttccgcgc acatttcccc gaaaagtgcc acctgacgcg ccctgtagcg gcgcattaag
     5761 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc
     5821 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     5881 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     5941 aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     6001 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     6061 actcaaccct atctcggtct attcttttga tttataaggg attttgccga tttcggccta
     6121 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac
     6181 g
//
